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Lab Log

LacZY plasmid

Keys to the table:
[?/?]: Names of restriction enzyme used for digestion:; E, EcoRI; X, XbaI; S, SpeI; P, PstI
- A ribosome binding site (RBS) is linked upstream to each gene
- The host cells used for transformation were competent JM109 E. coli cells

PCR amplification of the lacZ and lacY genes

Week 1 : May 18 – 22
Date	Group 1	Group 2	Group 3	Group 4
Monday 18	==introduction==
Tuesday 19
Wednesday 20	PCR amplification of the BBa_S04055 lacZ gene	PCR amplification of the BBa_S04055 lacZ gene	PCR amplification of the BBa_S04055 lacY gene	PCR amplification of the BBa_S04055 lacY gene
Thursday 21	Gel electrophoresis of the amplicon lacZ PCR product purification Restriction digestion with [E/P] of the vector pSB1C3 & lacZ	Gel electrophoresis of the amplicon lacZ PCR product purification Restriction digestion with [E/P] of the vector pSB1C3 & lacZ	Gel electrophoresis of the amplicon lacY PCR product purification Restriction digestion with [E/P] of the vector pSB1C3 & lacY	Gel electrophoresis of the amplicon lacY PCR product purification Restriction digestion with [E/P] of the vector pSB1C3 & lacY
Friday 22	Gel purification of vector pSB1C3 [E/P] digest Ligation of the [E/P] digested lacZ insert into the [E/P] digested vector pSB1C3	Gel purification of vector pSB1C3 [E/P] digest Ligation of the [E/P] digested lacZ insert into the [E/P] digested vector pSB1C3	Gel purification of vector pSB1C3 [E/P] digest Ligation of the [E/P] digested lacY insert into the [E/P] digested vector pSB1C3	Gel purification of vector pSB1C3 [E/P] digest Ligation of the [E/P] digested lacY insert into the [E/P] digested vector pSB1C3

Week 2 : May 25 – 29
Date	Group 1	Group 2	Group 3	Group 4
Monday 25
Tuesday 26	Transformation of May 22 ligation product into competent E. coli cells	Transformation of May 22 ligation product into competent E. coli cells	Transformation of May 22 ligation product into competent E. coli cells	Transformation of May 22 ligation product into competent E. coli cells
Wednesday 27	Colony PCR of May 26 transformed cells	Colony PCR of May 26 transformed cells	Colony PCR of May 26 transformed cells	Colony PCR of May 26 transformed cells
Thursday 28	Extraction of the plasmid pSB1C3-BBa_J23100 Restriction digestion with [S/P] of the vector pSB1C3-BBa_J23100	Extraction of the plasmid pSB1C3-BBa_J23100 Restriction digestion with [S/P] of the vector pSB1C3-BBa_J23100	Extraction of the plasmid pSB1C3-BBa_J23100 Restriction digestion with [S/P] of the vector pSB1C3-BBa_J23100	Extraction of the plasmid pSB1C3-BBa_J23100 Restriction digestion with [S/P] of the vector pSB1C3-BBa_J23100
Friday 29	Gel purification of May 28 digest	Gel purification of May 28 digest	Gel purification of May 28 digest	Gel purification of May 28 digest

*Groups 1 & 3 and Groups 2 & 4 used different cloning strategies to assemble the biobrick J23100_lacZ-lacY.

Week 3 : June 1 – 5
Date	Group 1	Group 3	Group 2	Group 4
Monday 1	Extraction of the plasmid pSB1C3-lacZ (from May 26 transformed cells) Restriction digestion with [X/P] of the insert lacZ Gel purification	Extraction of the plasmid pSB1C3-lacY (from May 26 transformed cells) Restriction digestion with [X/P] of the insert lacY Gel purification	Extraction of the plasmid pSB1C3-lacZ from Gp1 Restriction digestion with [E/S] of the insert lacZ Gel purification	Extraction of the plasmid pSB1C3-lacY from Gp1 Restriction digestion with [E/S] of the insert lacY Gel purification
Tuesday 2	Redo the work on June 1	Redo the work on June 1	Redo the work on June 1	Redo the work on June 1
Wednesday 3	Ligation of Gp1 [X/P] digested lacZ insert (from June 2) into [S/P] digested vector pSB1C3-BBa_J23100 (from May 29) Transformation		Ligation of Gp2 [E/S] digested lacZ insert (from June 2) into Gp4 [E/X] digested vector pSB1C3-lacY(from June 2) Transformation
Thursday 4	Colony PCR on June 3 transformed cells		Colony PCR on June 3 transformed cells
Friday 5	Extraction of plasmid pSB1C3-BBa_J23100-lacZ (from June 3 transformed cells) Restriction digestion with [S/P] of the vector pSB1C3-BBa_J23100-lacZ Gel purification		Extraction of plasmid pSB1C3-lacZ-lacY (from June 3 transformed cells) Restriction digestion with [X/P] of the insert lacZ-lacY Gel purification

Week 4 : June 8 – 12
Date	Group 1	Group 3	Group 2	Group 4
Monday 8	Ligation of Gp3 [X/P] digested lacY insert (from June 2) into [S/P] digested vector pSB1C3-BBa_J23100-lacZ (from June 5) Transformation		Ligation of [X/P] digested lacZ-lacY insert (from June 5) into [S/P] digested vector pSB1C3-BBa_J23100 (from May 29) Transformation
Tuesday 9	Colony PCR of June 8 transformed cells (failed)		Colony PCR of June 8 transformed cells (failed)
Wednesday 10	Extraction of the plasmid pSB1C3-BBa_J23100-lacZ-lacY (from June 8 transformed cells) Restriction analysis with [E/P] for confirmation of the insert size		Extraction of the plasmid pSB1C3-BBa_J23100-lacZ-lacY (from June 8 transformed cells) Restriction analysis with [E/P] for confirmation of the insert size

Characterization of the lacZY plasmid

Week 8 : July 6 – 10
Date		Remarks
Monday 6
Tuesday 7	RNA extraction from the E. coli cells harboring the recombinant plasmid pSB1C3-BBa_J23100- lacZ-lacY	Not successful
Wednesday 8	Redo RNA extraction	Not successful
Thursday 9
Friday 10	Redo RNA extraction	Not successful

Week 9 : July 13 – 18
Date		Remarks
Monday 13	Redo the RNA extraction	Successful
Tuesday 14	Extract RNA from control E. coli cells	Successful
Wednesday 15	Redo RNA extraction from control E. coli cells Perform RT-PCR on purified RNA from recombinant and control E. coli	Successful
Thursday 16	Check the size of the RT-PCR product using gel electrophoresis	Successful
Friday 17	Prepare Z-buffer
Saturday 18	Perform qPCR on the control cDNA	Successful

Week 10 : July 20 – 25
Date		Remarks
Monday 20	Perform q-PCR on recombinant cDNA	Successful
Tuesday 21	Perform ONPG assay -characterization on the expression level of LacZ protein
Wednesday 22
Thursday 23
Friday 24

Lysis plasmid (λ phage)

Keys to the table:
ori: original gene sequence, without codon optimization
co: codon optimized
λ Lysis(o_o_o): SλWT(ori)-Rλ(ori)-Rzλ(ori)
λLysis(c_c_c): SλWT(co)-Rλ(co)-Rzλ(co)
λLysis_Sm(o_c_c): Sλmut(ori)-Rλ(co)-Rzλ(co)
λLysis_Sm(c_c_c): Sλmut(co)-Rλ(co)-Rzλ(co)
[x/y]: restriction sites digested: E, EcoRI; X, XbaI; S, SpeI; P, PstI
Gene name or plasmid name [x/y]: the gene fragment or plasmid flanked with the restriction sites x and y at the ends
-A ribosome binding site (RBS) is linked upstream of each gene
-Coloring refers to sub-tasks in that particular group
-The cells used for transformation were competent JM109 E. coli cells

Week 4 : June 8 – 12

Date Group 1 Group 2 Group 3 Group 4

Monday 8

Tuesday 9

Wednesday 10
Extraction of the plasmid pGOv4-Rzλ(co)

Extraction of the plasmid pGOv4-Rλ(co)

Thursday 11
Preparation of the plasmid pSB1C3

Restriction digestion with [E/P]

Restriction digestion with [E/P] on the plasmid pGOv4-Rzλ(co)

Gel purification

Restriction digestion with [E/P] on the plasmid pGOv4-Rλ(co)

Gel purification

Friday 12
Ligation of the [E/P] digested Rzλ(co) insert (from June 11) with the [E/P] digested vector pSB1C3

Transformation of the ligaiton product into competent E. coli cells

Redo 11/6’s work

Week 5 : June 15 – 19

Date Group 1 Group 2 Group 3 Group 4

Monday 15

Tuesday 16
Colony PCR on June 12 transformed cells (Result: no bands amplified)

Ligation of the [E/P] digested Rλ(co) insert (from June 12) into the [E/P] digested vector pSB1C3

Transformation of the ligation product into competent E. coli cells

Wednesday 17
Colony PCR on June 16 transformed cells (Result: no bands amplified)

Thursday 18
Redo the ligation of the [E/P] digested Rzλ(co) insert into the [E/P] digested vector pSB1C3

Transformation of the ligation product into competent E. coli cells

Redo the extraction of the plasmid pGOv4-Rλ(co)

Restriction digestion with [E/P]

Friday 19
Colony PCR on June 18 transformed cells (Result: seen bands of the expected size)

Gel purification of June 18 digest

Week 6 : June 22 – 26

Date Group 1 Group 2 Group 3 Group 4

Monday 22

Tuesday 23
Redo the extraction of the plasmid pGOv4-Rλ(co)

Restriction digestion with [E/P]

Gel purification

Wednesday 24

Thursday 25
Ligation of the [E/P] digested Rλ(co) insert into the [E/P] digested pSB1C3 vector

Transformation of the ligation product into competent E. coli cells

Friday 26
Colony PCR on June 25 transformed cells (Result: showed no bands)

Week 7 : June 29 – July 3

Date Group 1 Group 2 Group 3 Group 4

Monday 29
Extraction of the plasmid pSB1C3-Rzλ(co) (from June 18 transformed cells)

Colony PCR on June 25 transformed cells (Result: again showed no bands)

Tuesday 30
Sequencing result confirmed the presence of Rλ(co) insert in pSB1C3

Wednesday 1

Thursday 2
PCR amplification of the BBa_K124017 SλWT(ori) gene

PCR amplification of the BBa_K124017 Rλ(ori) gene

Rλ(ori) PCR product purification

Transformation of pGOv4-SλWT(co) into competent E. coli cells

PCR amplification of the BBa_K124017 Rzλ(ori) gene

Rzλ(ori) PCR product purification

Restriction digestion with [E/P]

PCR amplification of the BBa_K124017 Rλ(ori) gene

Rλ(ori) PCR product purification

Friday 3
Restriction digestion of pSB1C3 vector with [E/X]

Restriction digestion of Rλ(co) with [E/P]

Gel purification
[1]
Gel purification of July 2 [E/P] digested Rzλ(ori)

Ligation of the [E/P] digested Rzλ(ori) insert into [E/P] digested pSB1C3 vector
[2]
Extraction of the plasmid pSB1C3-Rλ(co) (from June 25 transformed cells)

Restriction digestion with [S/P]

Gel purification
[3]
Restriction digestion of June 2 Rzλ(ori) with [X/P] & Heat kill

Ligation of [X/P] digested Rzλ(ori) insert into [S/P] digested pSB1C3-Rλ(co) vector

Extraction of the plasmid pSB1C3-Rλ(co) (from Gp3 June 25 transformed cells)

Week 8 : July 6 – July 10

Date Group 1 Group 2 Group 3 Group 4

Monday 6
Gel purification of the [X/P] digested pSB1C3 vector isolated on July 3

PCR amplification of the BBa_K124017 Rzλ(ori) gene using a F-primer with SacII restriction site

Transformation of the ligation product pSB1C3-Rλ(co)-Rzλ(ori) (from July 3 [3])

Restriction digestion of July 3 Rλ(co) with [S/P]
Gel purification

Restriction digestion of Rzλ(co) (from Gp2 June 29) with [X/P]
Gel purification

Tuesday 7
Gel electrophoresis of the SacII digested Rzλ(ori) PCR amplicon

PCR amplification of SλWT(ori) using a new primer pair

Extraction of the plasmid pGOv4-SλWT(co)
Restriction digestion with [X] on July 3 [P] digested Rλ(ori)

Colony PCR on July 6 Rλ(co)-Rzλ(ori) clones showed no bands

Redo July 3’s ligation of [X/P] digested Rzλ(ori) insert into [S/P] digested pSB1C3-Rλ(co) vector

Transformation

Ligation of the [X/P] digested Rzλ(co) insert into the [S/P] digested pSB1C3-Rλ(co) vector

Transformation

Wednesday 8
PCR purification of July 6 SacII Rzλ(ori) PCR product

Ligation of [X/P] digested Rλ(ori) into Gp1 July 6 [X/P]
Transformation of 7/7’s ligation product Rλ(ori) in pSB1C3

Colony PCR on July 7 Rλ(co)-Rzλ(ori) clones still showed no bands

Colony PCR on July 7 Rλ(co)-Rzλ(co) clones showed no bands

Thursday 9
Restriction digestion of SacII Rzλ(ori) with [SacII/P]
Restriction digestion of Gp2 July 7 SλWT(co) with [E/S]
Gel electrophoresis of July 7 SλWT(ori)
PCR purification of SλWT(ori)

Colony PCR on Rλ(ori) (from July 8 transformed cells)

Restriction digestion of July 7 SλWT(co) with [E/P]

Ligation of [X/P] July 3 digested Rzλ(ori) insert into [X/P] digested pSB1C3 vector

Transformation

Friday 10
Restriction digestion of SλWT(ori) with [X/P] & Gel purification
Ligation of [X/P] digested SλWT(ori) insert into July 6 [X/P] digested pSB1C3 vector
Transformation
Gel purification of July 9 [E/S] digested SλWT(co)

Gel purification of July 9 [E/P] digested SλWT(co)

Colony PCR on July 9 Rzλ(ori) clones showed no bands
Restriction analysis on July 7 transformed cells
Another redo of the ligation of July 3 [X/P] digested Rzλ(ori) insert into July 3 [S/P] digested pSB1C3-Rλ(co)
Transformation

Extraction of the plasmid pSB1C3-Rλ(co)-Rzλ(co) (from July 7 transformed cells)

Restriction analysis of Rλ(co)-Rzλ(co) showed bands of the expected size

Week 9 : July 13 – July 18

Date Group 1 Group 2 Group 3 Group 4

Monday 13
Colony PCR on SλWT(ori) clones showed bands of the expected size
Ligation of [E/S] digested SλWT(co) insert into pSB1C3 vector
Transformation

Extraction of the plasmid pSB1C3-Rλ(ori)
Restriction digestion with [X/P]
Gel purification

Colony PCR on 10/7 transformed cells pSB1C3-Rλ(co)-Rzλ(ori)

Tuesday 14
Colony PCR on SλWT(co) clones showed bands of the expected size

Restriction digestion of Gp2 Rλ(ori) with [X/SacII]

Restriction analysis of [X/P] digested Rλ(ori)
Restriction digestion of Rzλ(co) with [X/P]
Gel purification

Extraction of the plasmid pSB1C3-Rλ(co)-Rzλ(co)

Restriction digestion with [X/P]

Wednesday 15
Gel purification of July 14 [X/SacII] digested Rλ(ori)

Ligation between [E/S] digested SλWT(co), [X/SacII] digested Rλ(ori), [SacII/P] digested Rzλ(ori) and [E/P] digested pSB1C3 backbone
Transformation

Extraction of the plasmid pSB1C3-Rλ(ori)
Restriction digestion with [S/P]

Extraction of the plasmid pSB1C3-Rλ(co)
Restriction digestion of Rλ(co) with [E/S]
Restriction digestion of [E/X] digested Rzλ(ori) with CIP & Gel purification
Ligation of the [E/S] digested Rλ(co) insert into [E/X] digested pSB1C3-Rzλ(ori)
Transformation

Gel purification of July 14 [X/P] digested Rλ(co)-Rzλ(co)

Thursday 16
Colony PCR on SλWT(co)- Rλ(ori)-Rzλ(ori) showed bands believed to be the desired insert

Gel purification of July 15 [S/P] digested Rλ(ori)

Ligation of the [X/P] digested Rzλ(co) insert into [S/P] digestd pSB1C3-Rλ(ori) vector

Colony PCR on July 15 transformed cells

Redo the ligation of [E/S] digested Rλ(co) insert into [E/X] digested pSB1C3-Rzλ(ori)

Transformation

Friday 17
Transformation of July 16 ligation product pSB1C3-Rλ(ori)-Rzλ(co)

Colony PCR on 16/7 transformed cells
Restriction digestion of Sλ(co) with [E/S] & Gel purification
Redo the ligation of [E/S] digested Rλ(co) insert into [E/X] digested pSB1C3-Rzλ(ori)
Transformation

Saturday 18
Extraction of the plasmids pSB1C3-Sλmut(ori) and pSB1C3-Sλmut(co)
Restriction digestion with [S/P]

DNA sequencing results later revealed that the genes amplified from the BioBrick, BBa_K124017, did not contain the desired sequences. Genomic DNA from E. coli BL21(DE3) will be used as template (starting from week 10) to create new λ(ori) related BioBricks. Gene sequences of the λ(co) clones were correct.

Week 10 : July 20 – July 25

Date Group 1 Group 2 Group 3 Group 4

Monday 20
Gel purification of July 18 [S/P] digested pSB1C3-Sλmut(ori) and pSB1C3-Sλmut(co)
Ligation of July 15 [X/P] digested Rλ(co)-Rzλ(co) insert into the [S/P] digested pSB1C3-Sλmut(ori) vector
Ligation of July 15 [X/P] digested Rλ(co)-Rzλ(co) insert into the [S/P] digested pSB1C3-Sλmut(co) vector

Tuesday 21
Transformation of pGOv4-Ribo_Sλ(co), pGOv4-Ribo_Rλ(co) and pGOv4-Ribo_Rzλ(co) into competent E. coli cells
Transformation of pGOv4-SλWT(co)-Rλ(co)-Rzλ(co) (λLysis(c_c_c))

Transformation of the pSB1C3-Sλmut(ori)-Rλ(co)-Rzλ(co) (λLysis_Sm(o_c_c))

Transformation of the pSB1C3-Sλmut(co)-Rλ(co)-Rzλ(co) (λLysis_Sm(c_c_c))

Wednesday 22
Genomic DNA extraction of BL21(DE3) E. coli cells
PCR amplification of the Sλ(ori), Rλ(ori) and Rzλ(ori) genes using the extracted genomic DNA as template

Redo Gp1 July 21 transformation of the pGOv4-Ribo_Sλ(co) and pGOv4-SλWT(co)-Rλ(co)-Rzλ(co) (λLysis (c_c_c)) into competent E. coli cells

Colony PCR on λLysis_Sm(o_c_c) clones showed bands of the expected size

Colony PCR on c clones revealed that that the colonies contained inserts of the wrong sizes
Redo the restriction digestion of Rλ(co)-Rzλ(co) with [X/P] & Gel purification

Thursday 23
Extraction of the plasmids pGOv4-Ribo_Sλ(co), pGOv4-Ribo_Rλ(co) and pGOv4-Ribo_Rzλ(co)
Restriction digestion of the extracted plasmids with [E/P]
Gel purification

Extraction of the plasmid pGOv4-λLysis(c_c_c)

Restriction digestion with [E/P]

Redo the ligation of the [X/P] digested Rλ(co)-Rzλ(co) insert into the [S/P] digested pSB1C3-Sλmut(co) vector
Transformation

Friday 24
Ligation of the [E/P] digested Ribo_Sλ(co) and Ribo_Sλ(co) inserts into [E/P] digested pSB1C3 respectively
Transformation

Restriction digestion of the Sλ(ori), Rλ(ori) and Rzλ(ori) amplicons with [X/P]

Colony PCR on the new λLysis_Sm(c_c_c) clones showed bands of the expected size. However, later sequencing result revealed that the colony was actually a mixed one with both the correct plasmid and the self-ligated RRz(co) plasmid

Saturday 25
Colony PCR on July 24 transformed Ribo_Sλ and Ribo_Rλ clones

Ligation of the [X/P] digested Sλ(ori), Rλ(ori) and Rzλ(ori) inserts into [X/P] digested pSB1C3 respectively
Transformation

Week 11 : July 27 – August 1

Date Group 1 Group 2 Group 3 Group 4

Monday 27

Tuesday 28
Colony PCR on Sλ(ori), Rλ(ori) and Rzλ(ori) clones showed bands of the expected size

Restriction digestion of Ribo_Rλ(co) with [E/P]

Wednesday 29
Restriction digestion of λLysis_Sm(o_c_c) with [X/P]
Gel purification

Thursday 30

Friday 31 Redo the restriction digestion of Ribo_Rλ(co) with [E/P]
Redo the extraction of the plasmid pSB1C3-λLysis_Sm(o_c_c)
Redo the restriction digestion with [X/P]
Gel purification

Saturday 1
Extraction of the plasmid pSB1C3-BBa_J23100
Restriction digestion with [E/S] and [E/P] respectively & Gel purification
Restriction digestion of λLysis(c_c_c) with [X/P] & Gel purification

Week 12 : August 3 – August 9

Date Group 1 Group 2 Group 3 Group 4

Monday 3
Redo the ligation of the [X/P] digested Rλ(co)-Rzλ(co) insert into the [S/P] digested pSB1C3-Sλmut(co) vector

Extraction of the plasmid pGOv4-lacIq (pLacIQ_LacI_L8-UV5 lac)

Restriction digestion with [E/S]

Gel purification

Ligation of the [E/S] digested lacIq into [E/S] digested pSB1C3

Ligation of the [E/S] digested lacIq and [X/P] digested λLysis_Sm(o_c_c) into [E/P] digested pSB1C3

Tuesday 4
Transformation of Aug 3 ligation product

Wednesday 5
The Aug 4 transformed pSB1C3-lacIq showed no growth

Colony PCR on Aug 4 transformed λLysis_Sm(c_c_c) clones. The clones contained inserts of the correct size

Colony PCR on Aug 4 transformed lacIq-λLysis_Sm(o_c_c) clones. The clones contained inserts of the wrong sizes

Thursday 6
Restriction digestion of λLysis_Sm(o_c_c) with [E/X]

Gel purification

Redo the ligation of [E/S] digested lacIq into the [E/X] digested pSB1C3-λLysis_Sm(c_c_c)

Redo the ligation of the [E/S] digested lacIq into [E/S] digested pSB1C3

Friday 7
Redo the ligation of the [E/S] digested lacIq into the [E/X] digested pSB1C3-λLysis_Sm(o_c_c)
Transformation

Saturday 8
Restriction digestion of λLysis_Sm(c_c_c) with [E/X]

Gel purification

Sunday 9
Colony PCR on Aug 7 transformed cells. The lacIq clones contained inserts of the correct size. The lacIq-λLysis_Sm(o_c_c) clones showed no bands

Redo the restriction digestion of λLysis_Sm(c_c_c) with [E/X]

Gel purification

Ligation of the [E/S] digested lacIq into the [E/X] digested pSB1C3-λLysis_Sm(c_c_c)

Week 13 : August 10 – August 16

Date Group 1 Group 2 Group 3 Group 4

Monday 10
Redo the colony PCR on Aug 7 transformed lacIq-λLysis_Sm(o_c_c) cells. The clones contained inserts of the wrong sizes

Transformation of Aug 9 ligation product pSB1C3-lacIq-λLysis_Sm(c_c_c)

Lysis plasmid (phage 21)

Keys to the table:
ori: original gene sequence, without codon optimization
co: codon optimized
21Lysis(o_o_o):S21WT(ori)-Rλ(ori)-Rzλ(ori)
21Lysis(c_c_c):S21WT(co)-Rλ(co)-Rzλ(co)
21Lysis_Sm(o_c_c): S21mut(ori)-Rλ(co)-Rzλ(co)
21Lysis_Sm(c_c_c):S21mut(co)-Rλ(co)-Rzλ(co)
[x/y]: restriction sites digested: E, EcoRI; X, XbaI; S, SpeI; P, PstI
Gene name or plasmid name [x/y]: the gene fragment or plasmid flanked with the restriction sites x and y at the ends
-A ribosome binding site (RBS) is linked upstream of each gene
-Coloring refers to sub-tasks in that particular group
-The cells used for transformation were competent JM109 E. coli cells

Week 4 : June 8 – 12

Date Group 1 Group 2 Group 3 Group 4

Monday 8

Tuesday 9

Wednesday 10
Extraction of the plasmid pGOv4-Rz21(co) by mini-prep
Restriction digestion with [E/P]

Thursday 11
Gel purification of the June 10 [E/P] digested Rz21(co)
Ligation of the [E/P] digested Rz21(co) insert into [E/P] digested pSB1C3
Transformation

Friday 12
Colony PCR on Rz21(co) clones showed bands of the expected size

Week 5 : June 15 – 19

Date Group 1 Group 2 Group 3 Group 4

Monday 15

Tuesday 16
Plasmid extraction of the pGOv4-R21(co)
Restriction digestion with [E/P]

Wednesday 17
Gel purification of the [E/P] digested R21(co)
Ligation of the [E/P] digested R21(co) insert into the [E/P] digested pSB1C3
Transformation

Thursday 18
Colony PCR confirmed the presence of the R21(co) insert

Friday 19

Week 6 : June 22 – 26

Date Group 1 Group 2 Group 3 Group 4

Monday 22
Extraction of June 17 plasmid pSB1C3-R21(co)

Tuesday 23
Extraction of June 11 plasmid pSB1C3-Rz21(co)
Restriction analysis with [E/P]

Wednesday 24

Thursday 25

Friday 26
PCR amplification on phage 21 wild type lysis cassette

Week 7 : June 29 – July 3

Date Group 1 Group 2 Group 3 Group 4

Monday 29
Restriction digestion of June 22 R21(co) with [S/P] & Gel purification

Extraction of June 11 plasmid pSB1C3-Rz21(co)

Restriction digestion with [X/P]

Tuesday 30
Redo the PCR amplification on phage 21 wild type lysis cassette

Redo June 29’s restriction digestion of Rz21(co) with [X/P]

Wednesday 1

Thursday 2
Gel purification of [X/P] digested Rz21(co)

Ligation of the [X/P] digested Rz21(co) insert into Gp1 June 29 [S/P] digested pSB1C3-R21(co) in [S/P]

Friday 3
Transformation of July 2 ligation product pSB1C3-R21(co)-Rz21(co)

Week 8 : July 6 – July 10

Date Group 1 Group 2 Group 3 Group 4

Monday 6
Colony PCR on July 3 R21(co)-Rz21(co) clones. No bands were observed

Tuesday 7
Extraction of June 30 plasmid pSB1C3-S21WT(ori)-R21(ori)-Rz21(ori) (21Lysis(o_o_o))

Restriction digestion with [X/P]

Redo July 6's colony PCR with other colonies. No bands were observed

Wednesday 8
Redo the restriction digestion of 21Lysis(o_o_o) with [X/P]

Thursday 9
Gel purification of July 8 [X/P] digested 21Lysis(o_o_o)

Friday 10
Ligation of the [X/P] digested 21Lysis(o_o_o) insert into [X/P] digested pSB1C3 vector

Transformation

Restriction analysis on July 3 R21(co)-Rz21(co) clones. The analysis confirmed the presence of the R21(co)-Rz21(co) insert

Week 9 : July 13 – July 18

Date Group 1 Group 2 Group 3 Group 4

Monday 13
Colony PCR on July 10 21Lysis(o_o_o) clones. No bands were observed

Tuesday 14
Restriction analysis on the 21Lysis(o_o_o) clones. The analysis confirmed the presence of the 21Lysis(o_o_o) insert

Extraction of the plasmid pSB1C3-R21(co)-Rz21(co)

Restriction digestion with [X/P]

Wednesday 15
Gel purification of July 14 [X/P] digested R21(co)-Rz21(co)

Thursday 16

Friday 17

Saturday 18
Extraction of the plasmids pSB1C3-S21mut(ori) and pSB1C3-S21mut(co)

Restriction digestion wth [S/P]

Week 10 : July 20 – July 25

Date Group 1 Group 2 Group 3 Group 4

Monday 20
Gel purification of July 18 [S/P] digested pSB1C3-S21mut(ori) and pSB1C3-S21mut(co)

Ligation of July 15 [X/P] digested R21(co)-Rz21(co) insert into [S/P] digested pSB1C3-S21mut(ori)

Ligation of July 15 [X/P] digested R21(co)-Rz21(co) insert into [S/P] digested pSB1C3-S21mut(co)

Tuesday 21
Transformation of pGOv4-Ribo_S21WT(co), pGOv4-Ribo_R21(co) and pGOv4-Ribo_Rz21(co)

Transformation of July 20 ligation product pSB1C3-S21mut(ori)-R21(co)-Rz21(co) (21Lysis_Sm(o_c_c))

Transformation of July 20 ligation product pSB1C3-S21mut(co)-R21(co)-Rz21(co) (21Lysis_Sm(c_c_c))

Wednesday 22
Redo the transformation of pGOv4-Ribo_R21(co)

Colony PCR on the 21Lysis_Sm(o_c_c) clones revealed that the colonies contained inserts of the wrong sizes

Colony PCR on the 21Lysis_Sm(c_c_c) clones showed bands of the expected size

Redo the restriction digestion of R21(co)-Rz21(co) with [S/P]
Gel purification

Thursday 23
Extraction of the plasmids pGOv4-Ribo_S21WT(co), pGOv4-Ribo_R21(co) and pGOv4-Ribo_Rz21(co)
Restriction digestion of Ribo_S21WT(co) with [E/P] & [E/S], Ribo_R21(co) with [E/P] & [X/P] and Ribo_Rz21(co) with [E/P] & [X/P]

Redo the ligation of the [X/P] digested R21(co)-Rz21(co) insert into [S/P] digested pSB1C3-S21mut(ori)
Transformation
Ligation of [X/P] digested BBa_I13504 (GFP) insert into [S/P] digested 21Lysis(o_o_o)

Friday 24
Colony PCR on 21Lysis_Sm(o_c_c) clones showed bands of the expected size

Saturday 25

Week 11 : July 27 – August 1

Date Group 1 Group 2 Group 3 Group 4

Monday 27

Tuesday 28
Extraction of the plasmid pGOv4-Ribo_Rz21(co)
Redo the restriction digestion of Ribo_S21WT(co) and Ribo_R21(co) with [E/P]

Wednesday 29
Ligation of the [E/P] digested Ribo_S21WT(co) insert into the [E/P] digested pSB1C3 vector [E/P] & transformation

Ligation of the [E/P] digested Ribo_R21(co) insert into [E/P] digested pSB1C3
Extraction of the plasmids pGOv4-Ribo_S21WT(co), pGOv4-Ribo_R21(co) and pGOv4-Ribo_Rz21(co)
Restriction digestion of Ribo_S21WT(co) with [E/S], Ribo_R21(co) with [X/P] and Ribo_Rz21(co) with [E/P]

Gel purification

Redo the ligation of the [X/P] digested GFP insert into the [S/P] digested pSB1C3-21Lysis(o_o_o)
Transformation

Thursday 30
Restriction digestion of Ribo_R21(co) with [X/P]

Colony PCR on 21Lysis(o_o_o)-GFP clones showed bands of the expected size

Friday 31
Restriction digestion of 21Lysis_Sm(o_c_c) with [X/P]
Gel purification

Redo the extraction of the plasmid pSB1C3-21Lysis_Sm(c_c_c)
Restriction digestion of 21Lysis_Sm(c_c_c) with [X/P]
Gel purification

Saturday 1

Week 12 : August 3 – 8

Date Group 1 Group 2 Group 3 Group 4

Monday 3
Extraction of plasmid pGOV4-S21WT(co)/li>
Restriction digestion with [E/S]

Gel purification

Ligation of the [E/S] digested S21WT(co) and July 22 [X/P] digested R21(co)-Rz21(co) into July 22 [E/P] digested pSB1C3

Extraction of the plasmid pGOv4-lacIq (pLacIQ_LacI_L8-UV5 lac)

Restriction digestion with [E/S]

Gel purification

Ligation of the [E/S] digested lacIq and [X/P] digested 21Lysis_Sm(o_c_c) into [E/P] digested pSB1C3

Ligation of the [E/S] digested lacIq promoter cassette and [X/P] digested 21Lysis_Sm(c_c_c) into [E/P] digested pSB1C3

Tuesday 4
Transformation of Aug 3 ligation product

Wednesday 5
Colony PCR on Aug 4 transformed pSB1C3-21Lysis(c_c_c). The clones contained inserts of the expected size

Colony PCR on Aug 4 transformed pSB1C3-lacIq-21Lysis_Sm(o_c_c). The clones contained the wrong inserts

Colony PCR on Aug 4 transformed pSB1C3-lacIq-21Lysis_Sm(c_c_c). The clones contained the wrong inserts

Thursday 6
Restriction digestion of 21Lysis(c_c_c) with [E/X]

Gel purification

Ligation of the [E/S] digested lacIq into the [E/X] digested pSB1C3-21Lysis(c_c_c)

Restriction digestion of 21Lysis_Sm(o_c_c) and 21Lysis_Sm(c_c_c) with [E/X]

Gel purification

Redo the ligation of the [E/S] digested lacIq into the [E/X] digested pSB1C3-21Lysis_Sm(o_c_c) and pSB1C3-21Lysis_Sm(c_c_c) respectively

Friday 7
Transformation of Aug 6 ligation product

Redo the restriction digestion with [E/S] on the plasmid pGOv4-S21WT(co)
Gel purification

Saturday 8
* Colony PCR on Aug 7 transformed cells. The clones lacIq-21Lysis(c_c_c), lacIq-21Lysis_Sm(o_c_c) and lacIq-21Lysis_Sm(c_c_c) all contained inserts of the expected sizes

Ligation of the [E/S] digested S21WT(co) into the [E/S] digested pSB1C3
Transformation

Week 13 : August 10 – 16

Date Group 1 Group 2 Group 3 Group 4

Monday 10
Colony PCR on Aug 8 transformed cells. The S21WT(co) clones contained inset of the expected size

Interlab study

Keys to the table:

-[?/?]: Names of restriction enzyme used for digestion:; E, EcoRI; X, XbaI; S, SpeI; P, PstI

-The host cells used for transformation were competent JM109 E. coli cells

Week 8 : July 6 – 10

Monday 6
Transformation of pSB1C3-BBa_J23101, pSB1C3-BBa_J23106 and pSB1C3-BBa_J23117, and pSB1A2-BBa_I13504 into competent E. coli cells

Tuesday 7

Wednesday 8
Extraction of the plasmids pSB1C3-BBa_J23101, pSB1C3-BBa_J23106, pSB1C3-BBa_J23117 and pSB1A2-BBa_I13504 by mini-prep

Thursday 9

Friday 10

Week 9 : July 13 – 18

Monday 13
Transformation of plasmid pSB1C3-BBa_I13504 into competent E. coli cells

Tuesday 14
Restriction digestion with [S/P] of the plasmid pSB1C3-BBa_J23101

Restriction digestion with [S/P] of the plasmid pSB1C3-BBa_J23106

Restriction digestion with [S/P] of the plasmid pSB1C3-BBa_J23117

Redo the transformation of plasmids pSB1C3-BBa_I20270 and pSB1C3-BBa_R0040

Wednesday 15
Extraction of plasmid pSB1C3-BBa_I13504
Restriction digestion with [X/P] of the plasmid pSB1C3-BBa_I13504

Thursday 16
Extraction of plasmids pSB1C3-BBa_I20270 and pSB1C3-BBa_R0040

Gel purification of the [X/P] digested I13504

Gel purification of July 14 [S/P] digested vectors pSB1C3-BBa_J23101, pSB1C3-BBa_J23106 and pSB1C3-BBa_J23117

Friday 17
Ligation of the [X/P] digested BBa_I13504 insert into [S/P] digested vector pSB1C3-BBa_J23101
Ligation of the [X/P] digested BBa_I13504 insert into [S/P] digested vector pSB1C3-BBa_J23106
Ligation of the [X/P] digested BBa_I13504 insert into [S/P] digested vector pSB1C3-BBa_J23117
Transformation of the 3 ligation product

Saturday 18

Week 10 : July 20 – 25

Monday 20
Redo the transformation of all July 17 ligation product

Tuesday 21
Redo the transformation of July 17 pSB1C3-BBa_J23101-I13504 and pSB1C3-BBa_J23117-BBa_I13504 ligation product

Wednesday 22
Extraction of plasmid pSB1C3-BBa_J23106-I13504
Restriction analysis with [E/P] of the plasmid pSB1C3-BBa_J23106-BBa_I13504

Thursday 23
Restriction analysis with [E/P] of the plasmids pSB1C3-BBa_J23101-I13504 and pSB1C3-BBa_J23117-I13504

Friday 24

Saturday 25

Week 11 : July 27 – August 1

Monday 27

Tuesday 28
Checking of O.D A600, A511 - Positive control (I20270), negative control (R0040) and the J23101, J23106 and J23117

Redo the transformation of pSB1C3-BBa_J23117

Wednesday 29

Thursday 30
Extraction of the plasmid pSB1C3-BBa_J23117

Restriction digestion [S/P] of pSB1C3-BBa_J23117

Restriction digestion [X/P] of pSB1A2-BBa_I13504

Gel purification

Friday 31
Ligation of the [X/P] digested BBa_I13504 insert into the [S/P] digested pSB1C3-J23117
Transformation

Saturday 1
Extraction of the plasmids pSB1C3-BBa_J23117 and pSB1A2_BBa_I13504
Extraction of the plasmids pSB1C3-BBa_J23117 and pSB1A2_BBa_I13504

Week 12 : August 3 – 8

Monday 3
Redo the extraction of the plasmids pSB1C3-BBa_J23117 and pSB1A2_BBa_I13504
Restriction analysis [E/P]

Tuesday 4
Redo the ligation of the [X/P] digested BBa_I13504 into the [S/P] digested pSB1C3-BBa_J23117

ABOUT US

We are a diverse team of CityU undergraduates, working hard to create a better world.
read more

LOCATION

Department of Biology and Chemistry,
City University of Hong Kong
Tat Chee Avenue, Kowloon,
Hong Kong SAR

CONTACT US

Email: cityu.igem2015@gmail.com
Tel: +852 34427654

FOLLOW US ON

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-.wsite-background {background-image: url('https://static.igem.org/mediawiki/2015/f/f9/2015CityU_HK_notebook_banner.png') !important;background-repeat: no-repeat !important;background-position: 50% 82.7300033569336% !important;background-size: 100% !important;background-color: transparent !important;}
+.wsite-background {background-image: url('https://static.igem.org/mediawiki/2015/f/f9/2015CityU_HK_notebook_banner.png') !important;background-repeat: no-repeat !important;background-position: 50% 82.7300033569336% !important;background-size: 100% !important;background-color: transparent !}
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+#main-wrap{margin-left:auto; margin-right:auto; width:80%;}
+.wsite-headline {color:transparent;}
 </style>
 </head>
@@ Line 221: / Line 216: @@
 			<div class="container">
 				<div class="banner">
-					<h2 style="color:"#F5F5F5"><span class="wsite-text wsite-headline">
+					<h2><span class="wsite-text wsite-headline">
-	                                Notebook
+	                               Notebook
                                          </span></h2>
 				</div>
@@ Line 233: / Line 228: @@
              <ul>
                  <li>
-                     <a href="#laczy">
+                     <a  class="scroll" href="#laczy">
                        <i class="sidebar-fa"></i>
                          <span class="nav-text" id="first-item">
@@ Line 242: / Line 237: @@
                  <li class="has-subnav">
-                     <a href="#lysis-lambda">
+                     <a  class="scroll" href="#lysis-lambda">
                        <i class="sidebar-fa fa fa-th-list fa-2x"></i>
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+                     <a class="scroll" href="#lysis-21">
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-                     <a href="#interlab">
+                     <a  class="scroll" href="#interlab">
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@@ Line 283: / Line 278: @@
                     <center><h2>Lab Log</h2></center>
-<div class="wsite-spacer" style="height:50px;"></div>
+<div id="laczy" class="wsite-spacer" style="height:50px;"></div>
+<div><div style="height: 20px; overflow: hidden; width: 100%;"></div>
-<divid="laczy" ><div style="height: 20px; overflow: hidden; width: 100%;"></div>
 <hr class="styled-hr" style="width:100%;"></hr>
 <div style="height: 20px; overflow: hidden; width: 100%;"></div></div>
-<div class="paragraph" style="text-align:left;"><font size="5">lacZY plasmid</font></div></div>
+<div class="paragraph" style="text-align:left;"><font size="5">LacZY plasmid</font></div></div>
 <hr class="styled-hr" style="width:100%;"></hr>
 <div><div style="height: 20px; overflow: hidden; width: 100%;"></div>
-<p>Keys to the table: <br>
+<p><b>Keys to the table:</b> <br>
+<b>
 [?/?]: Names of restriction enzyme used for digestion:; E, EcoRI; X, XbaI; S, SpeI; P, PstI <br>
@@ Line 301: / Line 295: @@
 - The host cells used for transformation were competent JM109 <i>E. coli</i> cells </p>
+</b>
 <style>
 table {
@@ Line 324: / Line 318: @@
 <br>
-<p>PCR amplification of the lacZ and lacY genes.</p>
+<p><u><font size ="4"><center>PCR amplification of the lacZ and lacY genes</font></center></u></p>
 <table id="t01" border="1">
    <tr>
@@ Line 413: / Line 407: @@
 <br><br>
-<p>Groups 1 & 3 and Groups 2 & 4 used different cloning strategies to assemble the biobrick J23100_lacZ-lacY.</p>
+<p><font size="4">*Groups 1 & 3 and Groups 2 & 4 used different cloning strategies to assemble the biobrick J23100_lacZ-lacY.</font></p>
 <br>
 <table id="t01" border="1">
@@ Line 486: / Line 480: @@
 <br>
-<center><u>Characterization of the lacZY plasmid</u></center>
+<center><u><font size = "4">Characterization of the lacZY plasmid</font></u></center>
 <br>
@@ Line 603: / Line 597: @@
 </table>  <!-- end of lacZY -->
+<!---------lysis lambda ----->
+<div id="lysis-lambda" class="wsite-spacer" style="height:50px;"></div>
+<div><div style="height: 20px; overflow: hidden; width: 100%;"></div>
+<hr class="styled-hr" style="width:100%;"></hr>
+<div style="height: 20px; overflow: hidden; width: 100%;"></div></div>
+<div class="paragraph" style="text-align:left;"><font size="5">Lysis plasmid (λ phage)</font></div></div>
+<hr class="styled-hr" style="width:100%;"></hr>
+<div><div style="height: 20px; overflow: hidden; width: 100%;"></div>
+<b><p><b>Keys to the table:</b> <br>
+<b><u>ori:</u> original gene sequence, without codon optimization</b> <br>
+<b><u>co:</u> codon optimized </b><br>
+<b><u>λ Lysis(o_o_o)</u>: <i>SλWT(ori)-Rλ(ori)-Rzλ(ori)</i> </b><br>
+<b><u>λLysis(c_c_c)</u>: <i>SλWT(co)-Rλ(co)-Rzλ(co) </b><br></i>
+<b><u>λLysis_Sm(o_c_c)</u>: <i>Sλmut(ori)-Rλ(co)-Rzλ(co)</i></b> <br>
+<b><u>λLysis_Sm(c_c_c)</u>: <i>Sλmut(co)-Rλ(co)-Rzλ(co)</i> </b><br>
+<b>[x/y]: restriction sites digested: E, EcoRI; X, XbaI; S, SpeI; P, PstI <br>
+<b>Gene name or plasmid name [x/y]: the gene fragment or plasmid flanked with the restriction sites x and y at the ends <br>
+<b>-A ribosome binding site (RBS) is linked upstream of each gene <br>
+<b>-Coloring refers to sub-tasks in that particular group <br>
+<b>-The cells used for transformation were competent JM109 <i>E. coli</i> cells</p>
+</b>
+<style>
+table {
+    width:100%;
+}
+th, td {
+    padding: 5px;
+    text-align: left;
+}
+table#t01 tr:nth-child(even) {
+    background-color: #eee;
+}
+table#t01 tr:nth-child(odd) {
+   background-color:#fff;
+}
+table#t01 th	{
+    background-color: black;
+    color: white;
+}
+</style>
+<table id="t01" border="1">
+  <tr>
+    <th colspan="5">Week 4 : June 8 – 12</th>
+  </tr>
+  <tr>
+    <td width="20%">Date</td>
+    <td width="20%">Group 1</td>
+    <td width="20%">Group 2</td>
+    <td width="20%">Group 3</td>
+    <td width="20%">Group 4</td>
+  </tr>
+  <tr>
+    <td>Monday 8</td>
+    <td></td>
+    <td></td>
+    <td></td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Tuesday 9</td>
+    <td></td>
+    <td></td>
+    <td></td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Wednesday 10</td>
+    <td></td>
+    <td><li>Extraction of the plasmid pGOv4-<i>Rzλ(co)</i></li></td>
+    <td><li>Extraction of the plasmid pGOv4-<i>Rλ(co)</i></li></td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Thursday 11</td>
+    <td><li>Preparation of the plasmid pSB1C3</li> <li>Restriction digestion with [E/P]</li></td>
+    <td><li>Restriction digestion with [E/P] on the plasmid pGOv4-<i>Rzλ(co)</i> </li> <li>Gel purification</li></td>
+    <td><li>Restriction digestion with [E/P] on the plasmid pGOv4-<i>Rλ(co)</i> </li> <li>Gel purification</li></td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Friday 12</td>
+    <td></td>
+    <td><li>Ligation of the [E/P] digested <i>Rzλ(co)</i> insert (from June 11) with the [E/P] digested vector pSB1C3</li> <li>Transformation of the ligaiton product into competent <i>E. coli</i> cells</li></td>
+    <td><li>Redo 11/6’s work</li></td>
+    <td></td>
+  </tr>
+</table>
+<table id="t01" border="1">
+  <tr>
+    <th colspan="5">Week 5 : June 15 – 19</th>
+  </tr>
+  <tr>
+    <td width="20%">Date</td>
+    <td width="20%">Group 1</td>
+    <td width="20%">Group 2</td>
+    <td width="20%">Group 3</td>
+    <td width="20%">Group 4</td>
+  </tr>
+  <tr>
+    <td>Monday 15</td>
+    <td colspan="4" bgcolor="#FFB2D1"></td>
+  </tr>
+  <tr>
+    <td>Tuesday 16</td>
+    <td></td>
+    <td><li>Colony PCR on June 12 transformed cells (Result: no bands amplified)</li></td>
+    <td><li>Ligation of the [E/P] digested <i>Rλ(co)</i> insert (from June 12) into the [E/P] digested vector pSB1C3</li> <li>Transformation of the ligation product into competent <i>E. coli</i> cells</li></td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Wednesday 17</td>
+    <td></td>
+    <td></td>
+    <td><li>Colony PCR on June 16 transformed cells (Result: no bands amplified)</li></td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Thursday 18</td>
+    <td></td>
+    <td><li>Redo the ligation of the [E/P] digested <i>Rzλ(co)</i> insert into the [E/P] digested vector pSB1C3 </li> <li>Transformation of the ligation product into competent <i>E. coli</i> cells</li></td>
+    <td><li>Redo the extraction of the plasmid pGOv4-<i>Rλ(co)</i></li> <li>Restriction digestion with [E/P]</li></td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Friday 19</td>
+    <td></td>
+    <td><li>Colony PCR on June 18 transformed cells (Result: seen bands of the expected size)</li></td>
+    <td><li>Gel purification of June 18 digest</li></td>
+    <td></td>
+  </tr>
+</table>
+<table id="t01" border="1">
+  <tr>
+    <th colspan="5">Week 6 : June 22 – 26</th>
+  </tr>
+  <tr>
+    <td width="20%">Date</td>
+    <td width="20%">Group 1</td>
+    <td width="20%">Group 2</td>
+    <td width="20%">Group 3</td>
+    <td width="20%">Group 4</td>
+  </tr>
+  <tr>
+    <td>Monday 22</td>
+    <td></td>
+    <td></td>
+    <td></td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Tuesday 23</td>
+    <td></td>
+    <td></td>
+    <td><li>Redo the extraction of the plasmid pGOv4-<i>Rλ(co)</i></li> <li>Restriction digestion with [E/P]</li> <li>Gel purification</li></td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Wednesday 24</td>
+    <td></td>
+    <td></td>
+    <td></td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Thursday 25</td>
+    <td></td>
+    <td></td>
+    <td><li>Ligation of the [E/P] digested <i>Rλ(co)</i> insert into the [E/P] digested pSB1C3 vector</li> <li>Transformation of the ligation product into competent <i>E. coli</i> cells</li></td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Friday 26</td>
+    <td></td>
+    <td></td>
+    <td><li>Colony PCR on June 25 transformed cells (Result: showed no bands)</li></td>
+    <td></td>
+  </tr>
+</table>
+<table id="t01" border="1">
+  <tr>
+    <th colspan="5">Week 7 : June 29 – July 3</th>
+  </tr>
+  <tr>
+    <td width="20%">Date</td>
+    <td width="20%">Group 1</td>
+    <td width="20%">Group 2</td>
+    <td width="20%">Group 3</td>
+    <td width="20%">Group 4</td>
+  </tr>
+  <tr>
+    <td>Monday 29</td>
+    <td></td>
+    <td><li>Extraction of the plasmid pSB1C3-<i>Rzλ(co)</i> (from June 18 transformed cells)</li></td>
+    <td><li>Colony PCR on June 25 transformed cells (Result: again showed no bands)</li></td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Tuesday 30</td>
+    <td></td>
+    <td></td>
+    <td><li>Sequencing result confirmed the presence of <i>Rλ(co)</i> insert in pSB1C3</li></td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Wednesday 1</td>
+    <td colspan="4" bgcolor="#FFB2D1"></td>
+  </tr>
+  <tr>
+    <td>Thursday 2</td>
+    <td><li>PCR amplification of the BBa_K124017 <i>SλWT(ori)</i> gene</li></td>
+    <td><li><font color="#D633FF">PCR amplification of the BBa_K124017 <i>Rλ(ori)</i> gene</font></li> <li><font color="#D633FF"><i>Rλ(ori)</i> PCR product purification</font></li> <li><font color="#0099FF">Transformation of pGOv4-<i>SλWT(co)</i> into competent <i>E. coli</i> cells</font></li></td>
+    <td><li><font color="#D633FF">PCR amplification of the BBa_K124017 <i>Rzλ(ori)</i> gene</font></li> <li><font color="#D633FF"><i>Rzλ(ori)</i> PCR product purification</font></li> <li><font color="#D633FF">Restriction digestion with [E/P]</font></li></td>
+    <td><li>PCR amplification of the BBa_K124017 <i>Rλ(ori)</i> gene</li> <li><i>Rλ(ori)</i> PCR product purification</li></td>
+  </tr>
+  <tr>
+    <td>Friday 3</td>
+    <td><li>Restriction digestion of pSB1C3 vector with [E/X]</li></td>
+    <td><li>Restriction digestion of <i>Rλ(co)</i> with [E/P]</li> <li>Gel purification</li></td>
+    <td>[1]<li><font color="#D633FF">Gel purification of July 2 [E/P] digested <i>Rzλ(ori)</i></font></li> <li>Ligation of the [E/P] digested <font color="#D633FF"><i>Rzλ(ori)</i> insert </font>into [E/P] digested pSB1C3 vector</li> [2] <li><font color="#0099FF">Extraction of the plasmid pSB1C3-<i>Rλ(co)</i> (from June 25 transformed cells)</font></li> <li><font color="#0099FF">Restriction digestion with [S/P]</li> <li>Gel purification</font></li> [3] <li><font color="#D633FF">Restriction digestion of June 2 <i>Rzλ(ori)</i> with [X/P] &amp; Heat kill</font></li> <li><font color="#00CC00">Ligation of [X/P] digested Rzλ(ori) insert into [S/P] digested pSB1C3-<i>Rλ(co)</i> vector</li></td>
+    <td><li>Extraction of the plasmid pSB1C3-<i>Rλ(co)</i> (from Gp3 June 25 transformed cells)</li></td>
+  </tr>
+</table>
+<table id="t01" border="1">
+  <tr>
+    <th colspan="5">Week 8 : July 6 – July 10</th>
+  </tr>
+  <tr>
+    <td width="20%">Date</td>
+    <td width="20%">Group 1</td>
+    <td width="20%">Group 2</td>
+    <td width="20%">Group 3</td>
+    <td width="20%">Group 4</td>
+  </tr>
+  <tr>
+    <td>Monday 6</td>
+    <td><li>Gel purification of the [X/P] digested pSB1C3 vector isolated on July 3</li> <li><font color="#D633FF">PCR amplification of the BBa_K124017 <i>Rzλ(ori)</i> gene using a F-primer with SacII restriction site</font></li></td>
+    <td></td>
+    <td><li><font color="#00CC00">Transformation of the ligation product pSB1C3-<i>Rλ(co)-Rzλ(ori)</i> (from July 3 [3])</font></li></td>
+    <td><li>Restriction digestion of July 3 <i>Rλ(co)</i> with [S/P]</li><li>Gel purification</li> <br> <li>Restriction digestion of <i>Rzλ(co)</i> (from Gp2 June 29) with [X/P]</li><li>Gel purification</li></td>
+  </tr>
+  <tr>
+    <td>Tuesday 7</td>
+    <td><li><font color="#D633FF">Gel electrophoresis of the SacII digested <i>Rzλ(ori)</i> PCR amplicon</font></li> <li><font color="#0099FF">PCR amplification of <i>SλWT(ori)</i> using a new primer pair</font></li></td>
+    <td><li><font color="#0099FF">Extraction of the plasmid pGOv4-<i>SλWT(co)</i></font></li><li><font color="#D633FF">Restriction digestion with [X] on July 3 [P] digested <i>Rλ(ori)</i></font></li></td>
+    <td><li><font color="#00CC00">Colony PCR on July 6 <i>Rλ(co)-Rzλ(ori)</i> clones showed no bands</li> <li>Redo July 3’s ligation of [X/P] digested <i>Rzλ(ori)</i> insert into [S/P] digested pSB1C3-<i>Rλ(co)</i> vector</li> <li>Transformation</font></li></td>
+    <td><li>Ligation of the [X/P] digested <i>Rzλ(co)</i> insert into the [S/P] digested pSB1C3-<i>Rλ(co)</i> vector</li> <li>Transformation</li></td>
+  </tr>
+  <tr>
+    <td>Wednesday 8</td>
+    <td><li><font color="#D633FF">PCR purification of July 6 SacII <i>Rzλ(ori)</i> PCR product</font></li></td>
+    <td><li><font color="#D633FF">Ligation of [X/P] digested <i>Rλ(ori)</i> into Gp1 July 6 [X/P]</li><li>Transformation of 7/7’s ligation product <i>Rλ(ori)</i> in pSB1C3</font></li></td>
+    <td><li><font color="#00CC00">Colony PCR on July 7 <i>Rλ(co)-Rzλ(ori)</i> clones still showed no bands</font></li></td>
+    <td><li>Colony PCR on July 7 <i>Rλ(co)-Rzλ(co)</i> clones showed no bands</li></td>
+  </tr>
+  <tr>
+    <td>Thursday 9</td>
+    <td><li><font color="#D633FF">Restriction digestion of SacII <i>Rzλ(ori)</i> with [SacII/P]</font></li><li><font color="#00CC00">Restriction digestion of Gp2 July 7 <i>SλWT(co)</i> with [E/S]</font></li><li><font color="#0099FF">Gel electrophoresis of July 7 <i>SλWT(ori)</i></li><li>PCR purification of <i>SλWT(ori)</i></font></li></td>
+    <td><li><font color="#D633FF">Colony PCR on <i>Rλ(ori)</i> (from July 8 transformed cells)</font></li> <li><font color="#0099FF">Restriction digestion of July 7 <i>SλWT(co)</i> with [E/P]</font></li></td>
+    <td><li><font color="#D633FF">Ligation of [X/P] July 3 digested <i>Rzλ(ori)</i> insert into [X/P] digested pSB1C3 vector</li> <li>Transformation</font></li></td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Friday 10</td>
+    <td><li>Restriction digestion of <i>SλWT(ori)</i> with [X/P] &amp; Gel purification</li><li>Ligation of [X/P] digested <i>SλWT(ori)</i> insert into July 6 [X/P] digested pSB1C3 vector</li><li>Transformation</li><li>Gel purification of July 9 [E/S] digested <i>SλWT(co)</i></li></td>
+    <td><li><font color="#0099FF">Gel purification of July 9 [E/P] digested <i>SλWT(co)</i></font></li></td>
+    <td><li><font color="#D633FF">Colony PCR on July 9 <i>Rzλ(ori)</i> clones showed no bands</font></li><li><font color="#00CC00">Restriction analysis on July 7 transformed cells</li><li>Another redo of the ligation of July 3 [X/P] digested <i>Rzλ(ori)</i> insert into July 3 [S/P] digested pSB1C3-<i>Rλ(co)</i></li><li>Transformation</font></li></td>
+    <td><li>Extraction of the plasmid pSB1C3-<i>Rλ(co)-Rzλ(co)</i> (from July 7 transformed cells)</li> <li>Restriction analysis of <i>Rλ(co)-Rzλ(co)</i> showed bands of the expected size</li></td>
+  </tr>
+</table>
+<table id="t01" border="1">
+  <tr>
+    <th colspan="5">Week 9 : July 13 – July 18</th>
+  </tr>
+  <tr>
+    <td width="20%">Date</td>
+    <td width="20%">Group 1</td>
+    <td width="20%">Group 2</td>
+    <td width="20%">Group 3</td>
+    <td width="20%">Group 4</td>
+  </tr>
+  <tr>
+    <td>Monday 13</td>
+    <td><li><font color="#0099FF">Colony PCR on <i>SλWT(ori)</i> clones showed bands of the expected size</font></li><li><font color="#00CC00">Ligation of [E/S] digested <i>SλWT(co)</i> insert into pSB1C3 vector</li><li>Transformation</font></li></td>
+    <td><li><font color="#D633FF">Extraction of the plasmid pSB1C3-<i>Rλ(ori)</i></li><li>Restriction digestion with [X/P]</li><li>Gel purification </font></li></td>
+    <td><li><font color="#00CC00">Colony PCR on 10/7 transformed cells pSB1C3-<i>Rλ(co)-Rzλ(ori)</i></font></li></td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Tuesday 14</td>
+    <td><li><font color="#00CC00">Colony PCR on <i>SλWT(co)</i> clones showed bands of the expected size</font></li> <li><font color="#FF3333"> Restriction digestion of Gp2 <i>Rλ(ori)</i> with [X/SacII]</font></li></td>
+    <td><li><font color="#D633FF">Restriction analysis of [X/P] digested <i>Rλ(ori)</i></font></li><li>Restriction digestion of <i>Rzλ(co)</i> with [X/P]</li><li>Gel purification</li></td>
+    <td></td>
+    <td><li>Extraction of the plasmid pSB1C3-<i>Rλ(co)-Rzλ(co)</i></li> <li>Restriction digestion with [X/P]</li></td>
+  </tr>
+  <tr>
+    <td>Wednesday 15</td>
+    <td><li><font color="#FF3333">Gel purification of July 14 [X/SacII] digested <i>Rλ(ori)</i></font></li><br><li><font color="#FF9900">Ligation between</font> <font color="#00CC00">[E/S] digested <i>SλWT(co)</i></font>, <font color="#FF3333">[X/SacII] digested <i>Rλ(ori)</i></font>, <font color="#D633FF">[SacII/P] digested <i>Rzλ(ori)</i></font> and <font color="#FF9900">[E/P] digested pSB1C3 backbone</li><li>Transformation</font></li></td>
+    <td><li><font color="#D633FF">Extraction of the plasmid pSB1C3-<i>Rλ(ori)</i></li><li>Restriction digestion with [S/P]</font></li></td>
+    <td><li>Extraction of the plasmid pSB1C3-<i>Rλ(co)</i></li><li>Restriction digestion of <i>Rλ(co)</i> with [E/S]</li><li>Restriction digestion of [E/X] digested <i>Rzλ(ori)</i> with CIP &amp; Gel purification</li><li>Ligation of the [E/S] digested <i>Rλ(co)</i> insert into [E/X] digested pSB1C3-<i>Rzλ(ori)</i></li><li>Transformation</li></td>
+    <td><li>Gel purification of July 14 [X/P] digested <i>Rλ(co)-Rzλ(co)</i></li></td>
+  </tr>
+  <tr>
+    <td>Thursday 16</td>
+    <td><li><font color="#FF9900">Colony PCR on <i>SλWT(co)- Rλ(ori)-Rzλ(ori)</i> showed bands believed to be the desired insert</font></li></td>
+    <td><li><font color="#D633FF">Gel purification of July 15 [S/P] digested <i>Rλ(ori)</i></font></li> <li>Ligation of the [X/P] digested <i>Rzλ(co)</i> insert into <font color="#D633FF">[S/P] digestd pSB1C3-<i>Rλ(ori)</i> vector</font></li></td>
+    <td><li>Colony PCR on July 15 transformed cells</li> <li>Redo the ligation of [E/S] digested <i>Rλ(co)</i> insert into [E/X] digested pSB1C3-<i>Rzλ(ori)</i></li> <li>Transformation</li></td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Friday 17</td>
+    <td></td>
+    <td><li>Transformation of July 16 ligation product pSB1C3-<i>Rλ(ori)-Rzλ(co)</i></li></td>
+    <td><li>Colony PCR on 16/7 transformed cells</li><li>Restriction digestion of <i>Sλ(co)</i> with [E/S] &amp; Gel purification</li><li>Redo the ligation of [E/S] digested <i>Rλ(co)</i> insert into [E/X] digested pSB1C3-<i>Rzλ(ori)</i></li><li>Transformation</li></td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Saturday 18</td>
+    <td></td>
+    <td></td>
+    <td></td>
+    <td><li><font color="#D633FF">Extraction of the plasmids pSB1C3-<i>Sλmut(ori)</i> and pSB1C3-<i>Sλmut(co)</i></li><li>Restriction digestion with [S/P]</font></li></td>
+</table>
+<br>
+<font size="3">DNA sequencing results later revealed that the genes amplified from the BioBrick, BBa_K124017, did not contain the desired sequences. Genomic DNA from <i>E. coli</i> BL21(DE3) will be used as template (starting from week 10) to create new λ(ori) related BioBricks. Gene sequences of the λ(co) clones were correct.</font>
+<br><br>
+<table id="t01" border="1">
+  <tr>
+    <th colspan="5">Week 10 : July 20 – July 25</th>
+  </tr>
+  <tr>
+    <td width="20%">Date</td>
+    <td width="20%">Group 1</td>
+    <td width="20%">Group 2</td>
+    <td width="20%">Group 3</td>
+    <td width="20%">Group 4</td>
+  </tr>
+  <tr>
+    <td>Monday 20</td>
+    <td></td>
+    <td></td>
+    <td></td>
+    <td><li><font color="#D633FF"> Gel purification of July 18 [S/P] digested pSB1C3-<i>Sλmut(ori)</i> and pSB1C3-<i>Sλmut(co)</i></font></li><li><font color="#0099FF">Ligation of July 15 [X/P] digested <i>Rλ(co)-Rzλ(co)</i> insert into the [S/P] digested pSB1C3-<i>Sλmut(ori)</i> vector</font></li><li><font color="#FF9900"> Ligation of July 15 [X/P] digested <i>Rλ(co)-Rzλ(co)</i> insert into the [S/P] digested pSB1C3-<i>Sλmut(co)</i> vector</font></li></td>
+  </tr>
+  <tr>
+    <td>Tuesday 21</td>
+    <td></td>
+    <td><li>Transformation of pGOv4-Ribo_<i>Sλ(co)</i>, pGOv4-Ribo_<i>Rλ(co)</i> and pGOv4-Ribo_<i>Rzλ(co)</i> into competent <i>E. coli</i> cells</li><li>Transformation of pGOv4-<i>SλWT(co)-Rλ(co)-Rzλ(co)</i> (λLysis(c_c_c))</li></td>
+    <td></td>
+    <td><li><font color="#0099FF">Transformation of the pSB1C3-<i>Sλmut(ori)-Rλ(co)-Rzλ(co)</i> (λLysis_Sm(o_c_c))</font></li> <li><font color="#FF9900"> Transformation of the pSB1C3-<i>Sλmut(co)-Rλ(co)-Rzλ(co)</i> (λLysis_Sm(c_c_c))</font></li></td>
+  </tr>
+  <tr>
+    <td>Wednesday 22</td>
+    <td><li><font color=”#3333FF”>Genomic DNA extraction of BL21(DE3) <i>E. coli</i> cells</li><li>PCR amplification of the <i>Sλ(ori)</i>, <i>Rλ(ori)</i> and <i>Rzλ(ori)</i> genes using the extracted genomic DNA as template</font></li></td>
+    <td></td>
+    <td><li><font color="#FF00FF">Redo Gp1 July 21 transformation of the pGOv4-Ribo_<i>Sλ(co)</i> and pGOv4-<i>SλWT(co)-Rλ(co)-Rzλ(co)</i> (λLysis (c_c_c)) into competent <i>E. coli</i> cells</font></li></td>
+    <td><li><font color="#0099FF"> Colony PCR on λLysis_Sm(o_c_c) clones showed bands of the expected size</font></li> <br> <li><font color="#FF9900">Colony PCR on c clones revealed that that the colonies contained inserts of the wrong sizes</li><li>Redo the restriction digestion of <i>Rλ(co)-Rzλ(co)</i> with [X/P] &amp; Gel purification</font></li></td>
+  </tr>
+  <tr>
+    <td>Thursday 23</td>
+    <td><li><font color="#FF00FF">Extraction of the plasmids pGOv4-Ribo_<i>Sλ(co)</i>, pGOv4-Ribo_<i>Rλ(co)</i> and pGOv4-Ribo_<i>Rzλ(co)</i></li><li>Restriction digestion of the extracted plasmids with [E/P]</li><li>Gel purification</font></li></td>
+    <td></td>
+    <td><li><font color="#FF00FF">Extraction of the plasmid pGOv4-λLysis(c_c_c)</li> <li>Restriction digestion with [E/P]</font></li></td>
+    <td><li><font color="#FF9900">Redo the ligation of the [X/P] digested <i>Rλ(co)-Rzλ(co)</i> insert into the [S/P] digested pSB1C3-<i>Sλmut(co)</i> vector</li><li>Transformation</font></li></td>
+  </tr>
+  <tr>
+    <td>Friday 24</td>
+    <td><li><font color="#FF00FF">Ligation of the [E/P] digested Ribo_<i>Sλ(co)</i> and Ribo_<i>Sλ(co)</i> inserts into [E/P] digested pSB1C3 respectively</li><li>Transformation</font></li><br><li><font color="#3333FF">Restriction digestion of the <i>Sλ(ori)</i>, <i>Rλ(ori)</i> and <i>Rzλ(ori)</i> amplicons with [X/P]</font></li></td>
+    <td></td>
+    <td></td>
+    <td><li><font color="#FF9900">Colony PCR on the new λLysis_Sm(c_c_c) clones showed bands of the expected size. However, later sequencing result revealed that the colony was actually a mixed one with both the correct plasmid and the self-ligated <i>RRz(co)</i> plasmid</font></li></td>
+  </tr>
+  <tr>
+    <td>Saturday 25</td>
+    <td><li><font color="#FF00FF">Colony PCR on July 24 transformed Ribo_<i>Sλ</i> and Ribo_<i>Rλ</i> clones</font></li><br><li><font color="#3333FF">Ligation of the [X/P] digested <i>Sλ(ori)</i>, <i>Rλ(ori)</i> and <i>Rzλ(ori)</i> inserts into [X/P] digested pSB1C3 respectively</li><li>Transformation</font></li></td>
+    <td></td>
+    <td></td>
+    <td></td>
+</table>
+<table id="t01" border="1">
+<th colspan="5">Week 11 : July 27 – August 1</th>
+  </tr>
+  <tr>
+    <td width="20%">Date</td>
+    <td width="20%">Group 1</td>
+    <td width="20%">Group 2</td>
+    <td width="20%">Group 3</td>
+    <td width="20%">Group 4</td>
+  </tr>
+  <tr>
+    <td>Monday 27</td>
+    <td colspan="4" bgcolor="#FFB2D1"></td>
+  </tr>
+  <tr>
+    <td>Tuesday 28</td>
+    <td><li><font color="#3333FF">Colony PCR on <i>Sλ(ori)</i>, <i>Rλ(ori</i>) and <i>Rzλ(ori)</i> clones showed bands of the expected size</font></li></td>
+    <td></td>
+    <td><li>Restriction digestion of Ribo_<i>Rλ(co)</i> with [E/P]</li></td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Wednesday 29</td>
+    <td></td>
+    <td></td>
+    <td></td>
+    <td><li><font color="#0099FF">Restriction digestion of λLysis_Sm(o_c_c) with [X/P]</li><li>Gel purification</font></li></td>
+  </tr>
+  <tr>
+    <td>Thursday 30</td>
+    <td></td>
+    <td></td>
+    <td></td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Friday 31</td>
+    <td></td>
+    <td></td>
+    <td>Redo the restriction digestion of Ribo_<i>Rλ(co)</i> with [E/P]</td>
+    <td><li><font color="#0099FF"> Redo the extraction of the plasmid pSB1C3-λLysis_Sm(o_c_c)</li><li>Redo the restriction digestion with [X/P]</li><li>Gel purification</font></li></td>
+  </tr>
+  <tr>
+    <td>Saturday 1</td>
+    <td></td>
+    <td></td>
+    <td><li>Extraction of the plasmid pSB1C3-BBa_J23100</li><li>Restriction digestion with [E/S] and [E/P] respectively &amp; Gel purification</li><li><font color="#FF00FF">Restriction digestion of λLysis(c_c_c) with [X/P] &amp; Gel purification</font></li></td>
+    <td></td>
+</table>
+<table id="t01" border="1">
+<th colspan="5">Week 12 : August 3 – August 9</th>
+  </tr>
+  <tr>
+    <td width="20%">Date</td>
+    <td width="20%">Group 1</td>
+    <td width="20%">Group 2</td>
+    <td width="20%">Group 3</td>
+    <td width="20%">Group 4</td>
+  </tr>
+  <tr>
+    <td>Monday 3</td>
+    <td></td>
+    <td></td>
+    <td></td>
+    <td><li><font color="#FF9900">Redo the ligation of the [X/P] digested <i>Rλ(co)-Rzλ(co)</i> insert into the [S/P] digested pSB1C3-<i>Sλmut(co)</i> vector</font></li>
+<br>
+    <li><font color="#FF3333"> Extraction of the plasmid pGOv4-<i>lacIq</i> (pLacIQ_LacI_L8-UV5 lac)</font></li>
+    <li><font color="#FF3333">Restriction digestion with [E/S]</font></li>
+    <li><font color="#FF3333">Gel purification</font></li>
+    <li><font color="#FF3333">Ligation of the [E/S] digested <i>lacIq</i> into [E/S] digested pSB1C3</font></li>
+    <br><li><font color="#0099FF">Ligation of the [E/S] digested <i>lacIq</i> and [X/P] digested λLysis_Sm(o_c_c) into [E/P] digested pSB1C3</font></li></td>
+  </tr>
+  <tr>
+    <td>Tuesday 4</td>
+    <td></td>
+    <td></td>
+    <td></td>
+    <td><li>Transformation of Aug 3 ligation product</li></td>
+  </tr>
+  <tr>
+    <td>Wednesday 5</td>
+    <td></td>
+    <td></td>
+    <td></td>
+    <td><li><font color="#FF3333"> The Aug 4 transformed pSB1C3-<i>lacIq</i> showed no growth</font></li>
+    <li><font color="#FF9900">Colony PCR on Aug 4 transformed λLysis_Sm(c_c_c) clones. The clones contained inserts of the correct size</font></li>
+    <li><font color="#0099FF">Colony PCR on Aug 4 transformed <i>lacIq</i>-λLysis_Sm(o_c_c) clones. The clones contained inserts of the wrong sizes</font></li>
+    </td>
+  </tr>
+  <tr>
+    <td>Thursday 6</td>
+    <td></td>
+    <td></td>
+    <td></td>
+    <td><li><font color="#0099FF">Restriction digestion of λLysis_Sm(o_c_c) with [E/X]</li>
+    <li>Gel purification</font></li>
+    <li><font color="#FF9900">Redo the ligation of [E/S] digested <i>lacIq</i> into the [E/X] digested pSB1C3-λLysis_Sm(c_c_c)</font></li>
+    <li><font color="#FF3333"> Redo the ligation of the [E/S] digested <i>lacIq</i> into [E/S] digested pSB1C3</font></li></td>
+  </tr>
+  <tr>
+    <td>Friday 7</td>
+    <td></td>
+    <td></td>
+    <td></td>
+    <td><li><font color="#0099FF"> Redo the ligation of the [E/S] digested <i>lacIq</i> into the [E/X] digested pSB1C3-λLysis_Sm(o_c_c)</li><li>Transformation</font></li></td>
+  </tr>
+  <tr>
+    <td>Saturday 8</td>
+    <td></td>
+    <td></td>
+    <td></td>
+    <td><li><font color="#FF9900">Restriction digestion of λLysis_Sm(c_c_c) with [E/X]</li> <li>Gel purification</font></li></td>
+  <tr>
+    <td>Sunday 9</td>
+    <td></td>
+    <td></td>
+    <td></td>
+    <td><li>Colony PCR on Aug 7 transformed cells. <font color="#FF3333">The <i>lacIq</i> clones contained inserts of the correct size.</font> <font color="#0099FF"> The <i>lacIq</i>-λLysis_Sm(o_c_c) clones showed no bands</font></li>
+    <li><font color="#FF9900">Redo the restriction digestion of λLysis_Sm(c_c_c) with [E/X]</li>
+    <li>Gel purification</li>
+    <li>Ligation of the [E/S] digested <i>lacIq</i> into the [E/X] digested pSB1C3-λLysis_Sm(c_c_c)</font></li></td>
+</table>
+<table id="t01" border="1">
+<th colspan="5">Week 13 : August 10 – August 16</th>
+  </tr>
+  <tr>
+    <td width="20%">Date</td>
+    <td width="20%">Group 1</td>
+    <td width="20%">Group 2</td>
+    <td width="20%">Group 3</td>
+    <td width="20%">Group 4</td>
+  </tr>
+  <tr>
+    <td>Monday 10</td>
+    <td></td>
+    <td></td>
+    <td></td>
+    <td><li><font color="#0099FF">Redo the colony PCR on Aug 7 transformed <i>lacIq</i>-λLysis_Sm(o_c_c) cells. The clones contained inserts of the wrong sizes</font></li> <li><font color="#FF9900">Transformation of Aug 9 ligation product pSB1C3-<i>lacIq</i>-λLysis_Sm(c_c_c)</font></li></td>
+</table>
 <!---------lysis 21 --------->
-<div  id="lysis-21"><div style="height: 20px; overflow: hidden; width: 100%;"></div>
+<div id="lysis-21" class="wsite-spacer" style="height:50px;"></div>
+<div><div style="height: 20px; overflow: hidden; width: 100%;"></div>
 <hr class="styled-hr" style="width:100%;"></hr>
 <div style="height: 20px; overflow: hidden; width: 100%;"></div></div>
 <div class="paragraph" style="text-align:left;"><font size="5">Lysis plasmid (phage 21)</font></div></div>
@@ Line 616: / Line 1,176: @@
 <div><div style="height: 20px; overflow: hidden; width: 100%;"></div>
-<p>Keys to the table: <br>
+<p><b>Keys to the table: </b><br>
-<b>ori</b>: original gene sequence, without codon optimization <br>
+<b><u>ori</u>: original gene sequence, without codon optimization </b><br>
-<b>co</b>: codon optimized <br>
+<b><u>co</u>: codon optimized </b><br>
-<b>21Lysis(o_o_o):</b> <i>S21WT(ori)-Rλ(ori)-Rzλ(ori)</i><br>
+<b><u>21Lysis(o_o_o):</u><i>S21WT(ori)-Rλ(ori)-Rzλ(ori)</i></b><br>
-<b>21Lysis(c_c_c):</b> <i>S21WT(co)-Rλ(co)-Rzλ(co)</i><br>
+<b><u>21Lysis(c_c_c):</u><i>S21WT(co)-Rλ(co)-Rzλ(co)</i></b><br>
-<b>21Lysis_Sm(o_c_c):</b> <i>S21mut(ori)-Rλ(co)-Rzλ(co)</i><br>
+<b><u>21Lysis_Sm(o_c_c):</u> <i>S21mut(ori)-Rλ(co)-Rzλ(co)</i></b><br>
-<b>21Lysis_Sm(c_c_c):</b> <i>S21mut(co)-Rλ(co)-Rzλ(co)</i><br>
+<b><u>21Lysis_Sm(c_c_c):</u><i>S21mut(co)-Rλ(co)-Rzλ(co)</i></b><br>
-[x/y]: restriction sites digested: E, EcoRI; X, XbaI; S, SpeI; P, PstI <br>
+<b>[x/y]: restriction sites digested: E, EcoRI; X, XbaI; S, SpeI; P, PstI <br>
 Gene name or plasmid name [x/y]: the gene fragment or plasmid flanked with the restriction sites x and y at the ends <br>
@@ Line 638: / Line 1,198: @@
 -Coloring refers to sub-tasks in that particular group <br>
--The cells used for transformation were competent JM109 <i>E. coli</i> cells</p>
+-The cells used for transformation were competent JM109 <i>E. coli</i> cells</b></p>
 <style>
@@ Line 1,152: / Line 1,712: @@
 </table>  <!-- end of lysis 21 -->
+<!-------------------- Interlab ------------------->
+<div id="interlab" class="wsite-spacer" style="height:50px;"></div>
+<div><div style="height: 20px; overflow: hidden; width: 100%;"></div>
+<hr class="styled-hr" style="width:100%;"></hr>
+<div style="height: 20px; overflow: hidden; width: 100%;"></div></div>
+<div class="paragraph" style="text-align:left;"><font size="5">Interlab study</font></div></div>
+<hr class="styled-hr" style="width:100%;"></hr>
+<div><div style="height: 20px; overflow: hidden; width: 100%;"></div>
+<p><b>Keys to the table:</b> <br><br>
+<b>
+-[?/?]: Names of restriction enzyme used for digestion:; E, EcoRI; X, XbaI; S, SpeI; P, PstI <br><br>
+-The host cells used for transformation were competent JM109 <i>E. coli</i> cells</b></p><br>
+<style>
+table {
+    width:100%;
+}
+th, td {
+    padding: 5px;
+    text-align: left;
+}
+table#t01 tr:nth-child(even) {
+    background-color: #eee;
+}
+table#t01 tr:nth-child(odd) {
+   background-color:#fff;
+}
+table#t01 th	{
+    background-color: black;
+    color: white;
+}
+</style>
+<table id="t01" border="1">
+  <tr>
+    <th colspan="2">Week 8 : July 6 – 10</th>
+  </tr>
+  <tr>
+    <td>Monday 6</td>
+    <td><li>Transformation of pSB1C3-BBa_J23101, pSB1C3-BBa_J23106 and pSB1C3-BBa_J23117, and pSB1A2-BBa_I13504 into competent <i>E. coli</i> cells</li></td>
+  </tr>
+  <tr>
+    <td>Tuesday 7</td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Wednesday 8</td>
+    <td><li>Extraction of the plasmids pSB1C3-BBa_J23101, pSB1C3-BBa_J23106, pSB1C3-BBa_J23117 and pSB1A2-BBa_I13504 by mini-prep</li></td>
+  </tr>
+  <tr>
+    <td>Thursday 9</td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Friday 10</td>
+    <td></td>
+  </tr>
+</table>
+<table id="t01" border="1">
+  <tr>
+    <th colspan="2">Week 9 : July 13 – 18</th>
+  </tr>
+  <tr>
+    <td>Monday 13</td>
+    <td><li>Transformation of plasmid pSB1C3-BBa_I13504 into competent E. coli cells</li></td>
+  </tr>
+  <tr>
+    <td>Tuesday 14</td>
+    <td>
+    <li>Restriction digestion with [S/P] of the plasmid pSB1C3-BBa_J23101</li>
+    <li>Restriction digestion with [S/P] of the plasmid pSB1C3-BBa_J23106</li>
+    <li>Restriction digestion with [S/P] of the plasmid pSB1C3-BBa_J23117</li>
+    <li>Redo the transformation of plasmids pSB1C3-BBa_I20270 and pSB1C3-BBa_R0040</li>
+    </td>
+  </tr>
+  <tr>
+    <td>Wednesday 15</td>
+    <td><li>Extraction of plasmid pSB1C3-BBa_I13504</li><li>Restriction digestion with [X/P] of the plasmid pSB1C3-BBa_I13504</li></td>
+  </tr>
+  <tr>
+    <td>Thursday 16</td>
+    <td><li>Extraction of plasmids pSB1C3-BBa_I20270 and pSB1C3-BBa_R0040</li>
+    <li>Gel purification of the [X/P] digested I13504</li>
+    <li>Gel purification of July 14 [S/P] digested vectors pSB1C3-BBa_J23101, pSB1C3-BBa_J23106 and pSB1C3-BBa_J23117</li>
+    </td>
+  </tr>
+  <tr>
+    <td>Friday 17</td>
+    <td><li>Ligation of the [X/P] digested BBa_I13504 insert into [S/P] digested vector pSB1C3-BBa_J23101</li><li>Ligation of the [X/P] digested BBa_I13504 insert into [S/P] digested vector pSB1C3-BBa_J23106</li><li>Ligation of the [X/P] digested BBa_I13504 insert into [S/P] digested vector pSB1C3-BBa_J23117</li><li>Transformation of the 3 ligation product</li></td>
+  </tr>
+  <tr>
+    <td>Saturday 18</td>
+    <td></td>
+</tr>
+</table>
+<table id="t01" border="1">
+  <tr>
+    <th colspan="2">Week 10 : July 20 – 25</th>
+  </tr>
+  <tr>
+    <td>Monday 20</td>
+    <td><li>Redo the transformation of all July 17 ligation product</li></td>
+  </tr>
+  <tr>
+    <td>Tuesday 21</td>
+    <td>
+    <li>Redo the transformation of July 17 pSB1C3-BBa_J23101-I13504 and pSB1C3-BBa_J23117-BBa_I13504 ligation product</li>
+    </td>
+  </tr>
+  <tr>
+    <td>Wednesday 22</td>
+    <td><li>Extraction of plasmid pSB1C3-BBa_J23106-I13504</li><li>Restriction analysis with [E/P] of the plasmid pSB1C3-BBa_J23106-BBa_I13504</li></td>
+  </tr>
+  <tr>
+    <td>Thursday 23</td>
+    <td><li>Restriction analysis with [E/P] of the plasmids pSB1C3-BBa_J23101-I13504 and pSB1C3-BBa_J23117-I13504</li>
+    </td>
+  </tr>
+  <tr>
+    <td>Friday 24</td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Saturday 25</td>
+    <td></td>
+</tr>
+</table>
+<table id="t01" border="1">
+  <tr>
+    <th colspan="2">Week 11 : July 27 – August 1</th>
+  </tr>
+  <tr>
+    <td>Monday 27</td>
+    <td bgcolor="#FFB2D1"></td>
+  </tr>
+  <tr>
+    <td>Tuesday 28</td>
+    <td>
+    <li>Checking of O.D A600, A511 - Positive control (I20270), negative control (R0040) and the J23101, J23106 and J23117</li>
+    <li>Redo the transformation of pSB1C3-BBa_J23117</li>
+    </td>
+  </tr>
+  <tr>
+    <td>Wednesday 29</td>
+    <td></td>
+  </tr>
+  <tr>
+    <td>Thursday 30</td>
+    <td><li>Extraction of the plasmid pSB1C3-BBa_J23117</li> <li>Restriction digestion [S/P] of pSB1C3-BBa_J23117</li> <li>Restriction digestion [X/P] of pSB1A2-BBa_I13504</li> <li>Gel purification</li>
+    </td>
+  </tr>
+  <tr>
+    <td>Friday 31</td>
+    <td><li>Ligation of the [X/P] digested BBa_I13504 insert into the [S/P] digested pSB1C3-J23117</li><li>Transformation</li></td>
+  </tr>
+  <tr>
+    <td>Saturday 1</td>
+    <td><li>Extraction of the plasmids pSB1C3-BBa_J23117 and pSB1A2_BBa_I13504</li><li>Extraction of the plasmids pSB1C3-BBa_J23117 and pSB1A2_BBa_I13504</li></td>
+</tr>
+</table>
+<table id="t01" border="1">
+  <tr>
+    <th colspan="2">Week 12 : August 3 – 8</th>
+  </tr>
+  <tr>
+    <td>Monday 3</td>
+    <td><li>Redo the extraction of the plasmids pSB1C3-BBa_J23117 and pSB1A2_BBa_I13504</li><li>Restriction analysis [E/P]</li></td>
+  </tr>
+  <tr>
+    <td>Tuesday 4</td>
+    <td>
+    <li>Redo the ligation of the [X/P] digested BBa_I13504 into the [S/P] digested pSB1C3-BBa_J23117</li>
+    </td>
+  </tr>
+</table>  <!-- end of Interlay study -->
 			</div><!-- end container -->
-		</div><!-- end main-wrap -->
+		</div></div><!-- end main-wrap -->
@@ Line 1,190: / Line 1,943: @@
 		});
 	});
+       //smooth scrolling
+       $('.scroll').click(function() {
+        $('html,body').animate({
+           scrollTop:$($.attr(this,'href')).offset().top},500);
+            return false;
+        });
 });

Week 8 : July 6 – 10
Monday 6	Transformation of pSB1C3-BBa_J23101, pSB1C3-BBa_J23106 and pSB1C3-BBa_J23117, and pSB1A2-BBa_I13504 into competent E. coli cells
Tuesday 7
Wednesday 8	Extraction of the plasmids pSB1C3-BBa_J23101, pSB1C3-BBa_J23106, pSB1C3-BBa_J23117 and pSB1A2-BBa_I13504 by mini-prep
Thursday 9
Friday 10

Week 9 : July 13 – 18
Monday 13	Transformation of plasmid pSB1C3-BBa_I13504 into competent E. coli cells
Tuesday 14	Restriction digestion with [S/P] of the plasmid pSB1C3-BBa_J23101 Restriction digestion with [S/P] of the plasmid pSB1C3-BBa_J23106 Restriction digestion with [S/P] of the plasmid pSB1C3-BBa_J23117 Redo the transformation of plasmids pSB1C3-BBa_I20270 and pSB1C3-BBa_R0040
Wednesday 15	Extraction of plasmid pSB1C3-BBa_I13504 Restriction digestion with [X/P] of the plasmid pSB1C3-BBa_I13504
Thursday 16	Extraction of plasmids pSB1C3-BBa_I20270 and pSB1C3-BBa_R0040 Gel purification of the [X/P] digested I13504 Gel purification of July 14 [S/P] digested vectors pSB1C3-BBa_J23101, pSB1C3-BBa_J23106 and pSB1C3-BBa_J23117
Friday 17	Ligation of the [X/P] digested BBa_I13504 insert into [S/P] digested vector pSB1C3-BBa_J23101 Ligation of the [X/P] digested BBa_I13504 insert into [S/P] digested vector pSB1C3-BBa_J23106 Ligation of the [X/P] digested BBa_I13504 insert into [S/P] digested vector pSB1C3-BBa_J23117 Transformation of the 3 ligation product
Saturday 18

Week 10 : July 20 – 25
Monday 20	Redo the transformation of all July 17 ligation product
Tuesday 21	Redo the transformation of July 17 pSB1C3-BBa_J23101-I13504 and pSB1C3-BBa_J23117-BBa_I13504 ligation product
Wednesday 22	Extraction of plasmid pSB1C3-BBa_J23106-I13504 Restriction analysis with [E/P] of the plasmid pSB1C3-BBa_J23106-BBa_I13504
Thursday 23	Restriction analysis with [E/P] of the plasmids pSB1C3-BBa_J23101-I13504 and pSB1C3-BBa_J23117-I13504
Friday 24
Saturday 25

Week 11 : July 27 – August 1
Monday 27
Tuesday 28	Checking of O.D A600, A511 - Positive control (I20270), negative control (R0040) and the J23101, J23106 and J23117 Redo the transformation of pSB1C3-BBa_J23117
Wednesday 29
Thursday 30	Extraction of the plasmid pSB1C3-BBa_J23117 Restriction digestion [S/P] of pSB1C3-BBa_J23117 Restriction digestion [X/P] of pSB1A2-BBa_I13504 Gel purification
Friday 31	Ligation of the [X/P] digested BBa_I13504 insert into the [S/P] digested pSB1C3-J23117 Transformation
Saturday 1	Extraction of the plasmids pSB1C3-BBa_J23117 and pSB1A2_BBa_I13504 Extraction of the plasmids pSB1C3-BBa_J23117 and pSB1A2_BBa_I13504

Week 12 : August 3 – 8
Monday 3	Redo the extraction of the plasmids pSB1C3-BBa_J23117 and pSB1A2_BBa_I13504 Restriction analysis [E/P]
Tuesday 4	Redo the ligation of the [X/P] digested BBa_I13504 into the [S/P] digested pSB1C3-BBa_J23117