Difference between revisions of "Team:CityU HK/Notebook/Lysis plasmid (λ phage)"

Latest revision as of 03:43, 20 August 2015

Notebook - City University of Hong Kong 2015

Notebook

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)

@@ Line 346: / Line 346: @@
 <b>co</b>: codon optimized <br>
-<b>λ Lysis(o_o_o)</b>: SλWT(ori)-Rλ(ori)-Rzλ(ori) <br>
+<b>λ Lysis(o_o_o)</b>: <i>SλWT(ori)-Rλ(ori)-Rzλ(ori)</i> <br>
-<b>λLysis(c_c_c)</b>: SλWT(co)-Rλ(co)-Rzλ(co) <br>
+<b>λLysis(c_c_c)</b>: <i>SλWT(co)-Rλ(co)-Rzλ(co) <br></i>
-<b>λLysis_Sm(o_c_c)</b>: Sλmut(ori)-Rλ(co)-Rzλ(co) <br>
+<b>λLysis_Sm(o_c_c)</b>: <i>Sλmut(ori)-Rλ(co)-Rzλ(co)</i> <br>
-<b>λLysis_Sm(c_c_c)</b>: Sλmut(co)-Rλ(co)-Rzλ(co) <br>
+<b>λLysis_Sm(c_c_c)</b>: <i>Sλmut(co)-Rλ(co)-Rzλ(co)</i> <br>
 [x/y]: restriction sites digested: E, EcoRI; X, XbaI; S, SpeI; P, PstI <br>
@@ Line 388: / Line 388: @@
    <tr>
      <th colspan="5">Week 4 : June 8 – 12</th>
-  </tr>
-  <tr>
-    <td colspan="5"></td>
    </tr>
    <tr>
@@ Line 417: / Line 414: @@
      <td>Wednesday 10</td>
      <td></td>
-     <td><li>Extraction of the plasmid pGOv4-Rzλ(co)</li></td>
+     <td><li>Extraction of the plasmid pGOv4-<i>Rzλ(co)</i></li></td>
-     <td><li>Extraction of the plasmid pGOv4-Rλ(co)</li></td>
+     <td><li>Extraction of the plasmid pGOv4-<i>Rλ(co)</i></li></td>
      <td></td>
    </tr>
@@ Line 424: / Line 421: @@
      <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-Rzλ(co) </li> <li>Gel purification</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-Rλ(co) </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>
@@ Line 431: / Line 428: @@
      <td>Friday 12</td>
      <td></td>
-     <td><li>Ligation of the [E/P] digested Rzλ(co) 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>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></td>
+     <td><li>Redo 11/6’s work</li></td>
      <td></td>
    </tr>
@@ Line 440: / Line 437: @@
    <tr>
      <th colspan="5">Week 5 : June 15 – 19</th>
-  </tr>
-  <tr>
-    <td colspan="5"></td>
    </tr>
    <tr>
@@ Line 459: / Line 453: @@
      <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 Rλ(co) 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><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>
@@ Line 473: / Line 467: @@
      <td>Thursday 18</td>
      <td></td>
-     <td><li>Redo the ligation of the [E/P] digested Rzλ(co) 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 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-Rλ(co)</li> <li>Restriction digestion with [E/P]</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>
@@ Line 489: / Line 483: @@
    <tr>
      <th colspan="5">Week 6 : June 22 – 26</th>
-  </tr>
-  <tr>
-    <td colspan="5"></td>
    </tr>
    <tr>
@@ Line 511: / Line 502: @@
      <td></td>
      <td></td>
-     <td><li>Redo the extraction of the plasmid pGOv4-Rλ(co)</li> <li>Restriction digestion with [E/P]</li> <li>Gel purification</li></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>
@@ Line 526: / Line 517: @@
      <td></td>
      <td></td>
-     <td><li>Ligation of the [E/P] digested Rλ(co) 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><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>
@@ Line 541: / Line 532: @@
    <tr>
      <th colspan="5">Week 7 : June 29 – July 3</th>
-  </tr>
-  <tr>
-    <td colspan="5"></td>
    </tr>
    <tr>
@@ Line 555: / Line 543: @@
      <td>Monday 29</td>
      <td></td>
-     <td><li>Extraction of the plasmid pSB1C3-Rzλ(co) (from June 18 transformed cells)</li></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>
@@ Line 563: / Line 551: @@
      <td></td>
      <td></td>
-     <td><li>Sequencing result confirmed the presence of Rλ(co) insert in pSB1C3</li></td>
+     <td><li>Sequencing result confirmed the presence of <i>Rλ(co)</i> insert in pSB1C3</li></td>
      <td></td>
    </tr>
@@ Line 574: / Line 562: @@
    <tr>
      <td>Thursday 2</td>
-     <td><li>PCR amplification of the BBa_K124017 SλWT(ori) gene</li></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 Rλ(ori) gene</font></li> <li><font color="#D633FF">Rλ(ori) PCR product purification</font></li> <li><font color="#0099FF">Transformation of pGOv4-SλWT(co) into competent <i>E. coli</i> cells</font></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 Rzλ(ori) gene</font></li> <li><font color="#D633FF">Rzλ(ori) PCR product purification</font></li> <li><font color="#D633FF">Restriction digestion with [E/P]</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 Rλ(ori) gene</li> <li>Rλ(ori) PCR product purification</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 Rλ(co) with [E/P]</li> <li>Gel purification</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 Rzλ(ori)</font></li> <li>Ligation of the [E/P] digested <font color="#D633FF">Rzλ(ori) insert </font>into [E/P] digested pSB1C3 vector</li> [2] <li><font color="#0099FF">Extraction of the plasmid pSB1C3-Rλ(co) (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 Rzλ(ori) with [X/P] & Heat kill</font></li> <li><font color="#00CC00">Ligation of [X/P] digested Rzλ(ori) insert into [S/P] digested pSB1C3-Rλ(co) vector</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] & 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-Rλ(co) (from Gp3 June 25 transformed cells)</li></td>
+     <td><li>Extraction of the plasmid pSB1C3-<i>Rλ(co)</i> (from Gp3 June 25 transformed cells)</li></td>
    </tr>
 </table>
@@ Line 591: / Line 579: @@
    <tr>
      <th colspan="5">Week 8 : July 6 – July 10</th>
-  </tr>
-  <tr>
-    <td colspan="5"></td>
    </tr>
    <tr>
@@ Line 604: / Line 589: @@
    <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 Rzλ(ori) gene using a F-primer with SacII restriction site</font></li></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-Rλ(co)-Rzλ(ori) (from July 3 [3])</font></li></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 Rλ(co) with [S/P]</li><li>Gel purification</li> <br> <li>Restriction digestion of Rzλ(co) (from Gp2 June 29) with [X/P]</li><li>Gel purification</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 Rzλ(ori) PCR amplicon</font></li> <li><font color="#0099FF">PCR amplification of SλWT(ori) using a new primer pair</font></li></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-SλWT(co)</font></li><li><font color="#D633FF">Restriction digestion with [X] on July 3 [P] digested Rλ(ori)</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 Rλ(co)-Rzλ(ori) clones showed no bands</li> <li>Redo July 3’s ligation of [X/P] digested Rzλ(ori) insert into [S/P] digested pSB1C3-Rλ(co) vector</li> <li>Transformation</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 Rzλ(co) insert into the [S/P] digested pSB1C3-Rλ(co) vector</li> <li>Transformation</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 Rzλ(ori) PCR product</font></li></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 Rλ(ori) into Gp1 July 6 [X/P]</li><li>Transformation of 7/7’s ligation product Rλ(ori) in pSB1C3</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 Rλ(co)-Rzλ(ori) clones still showed no bands</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 Rλ(co)-Rzλ(co) clones showed no bands</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 Rzλ(ori) with [SacII/P]</font></li><li><font color="#00CC00">Restriction digestion of Gp2 July 7 SλWT(co) with [E/S]</font></li><li><font color="#0099FF">Gel electrophoresis of July 7 SλWT(ori)</li><li>PCR purification of SλWT(ori)</font></li></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 Rλ(ori) (from July 8 transformed cells)</font></li> <li><font color="#0099FF">Restriction digestion of July 7 SλWT(co) with [E/P]</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 Rzλ(ori) insert into [X/P] digested pSB1C3 vector</li> <li>Transformation</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 SλWT(ori) with [X/P] & Gel purification</li><li>Ligation of [X/P] digested SλWT(ori) insert into July 6 [X/P] digested pSB1C3 vector</li><li>Transformation</li><li>Gel purification of July 9 [E/S] digested SλWT(co)</li></td>
+     <td><li>Restriction digestion of <i>SλWT(ori)</i> with [X/P] & 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 SλWT(co)</font></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 Rzλ(ori) 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 Rzλ(ori) insert into July 3 [S/P] digested pSB1C3-Rλ(co)</li><li>Transformation</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-Rλ(co)-Rzλ(co) (from July 7 transformed cells)</li> <li>Restriction analysis of Rλ(co)-Rzλ(co) showed bands of the expected size</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>
@@ Line 644: / Line 629: @@
    <tr>
      <th colspan="5">Week 9 : July 13 – July 18</th>
-  </tr>
-  <tr>
-    <td colspan="5"></td>
    </tr>
    <tr>
@@ Line 657: / Line 639: @@
    <tr>
      <td>Monday 13</td>
-     <td><li><font color="#0099FF">Colony PCR on SλWT(ori) clones showed bands of the expected size</font></li><li><font color="#00CC00">Ligation of [E/S] digested SλWT(co) insert into pSB1C3 vector</li><li>Transformation</font></li></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-Rλ(ori)</li><li>Restriction digestion with [X/P]</li><li>Gel purification </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-Rλ(co)-Rzλ(ori)</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 SλWT(co) clones showed bands of the expected size</font></li> <li><font color="#FF3333"> Restriction digestion of Gp2 Rλ(ori) with [X/SacII]</font></li></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 Rλ(ori)</font></li><li>Restriction digestion of Rzλ(co) with [X/P]</li><li>Gel purification</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-Rλ(co)-Rzλ(co)</li> <li>Restriction digestion with [X/P]</li></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 Rλ(ori)</font></li><br><li><font color="#FF9900">Ligation between</font> <font color="#00CC00">[E/S] digested SλWT(co)</font>, <font color="#FF3333">[X/SacII] digested Rλ(ori)</font>, <font color="#D633FF">[SacII/P] digested Rzλ(ori)</font> and <font color="#FF9900">[E/P] digested pSB1C3 backbone</li><li>Transformation</font></li></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-Rλ(ori)</li><li>Restriction digestion with [S/P]</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-Rλ(co)</li><li>Restriction digestion of Rλ(co) with [E/S]</li><li>Restriction digestion of [E/X] digested Rzλ(ori) with CIP & Gel purification</li><li>Ligation of the [E/S] digested Rλ(co) insert into [E/X] digested pSB1C3-Rzλ(ori)</li><li>Transformation</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 & 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 Rλ(co)-Rzλ(co)</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 SλWT(co)- Rλ(ori)-Rzλ(ori) showed bands believed to be the desired insert</font></li></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 Rλ(ori)</font></li> <li>Ligation of the [X/P] digested Rzλ(co) insert into <font color="#D633FF">[S/P] digestd pSB1C3-Rλ(ori) vector</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 Rλ(co) insert into [E/X] digested pSB1C3-Rzλ(ori)</li> <li>Transformation</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>
@@ Line 688: / Line 670: @@
      <td>Friday 17</td>
      <td></td>
-     <td><li>Transformation of July 16 ligation product pSB1C3-Rλ(ori)-Rzλ(co)</li></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 Sλ(co) with [E/S] & Gel purification</li><li>Redo the ligation of [E/S] digested Rλ(co) insert into [E/X] digested pSB1C3-Rzλ(ori)</li><li>Transformation</li></td>
+     <td><li>Colony PCR on 16/7 transformed cells</li><li>Restriction digestion of <i>Sλ(co)</i> with [E/S] & 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>
@@ Line 698: / Line 680: @@
      <td></td>
      <td></td>
-     <td><li><font color="#D633FF">Extraction of the plasmids pSB1C3-Sλmut(ori) and pSB1C3-Sλmut(co)</li><li>Restriction digestion with [S/P]</font></li></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>
-<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.</p>
+<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><br>
 <table id="t01" border="1">
    <tr>
      <th colspan="5">Week 10 : July 20 – July 25</th>
-  </tr>
-  <tr>
-    <td colspan="5"></td>
    </tr>
    <tr>
@@ Line 724: / Line 703: @@
      <td></td>
      <td></td>
-     <td><li><font color="#D633FF"> Gel purification of July 18 [S/P] digested pSB1C3-Sλmut(ori) and pSB1C3-Sλmut(co)</font></li><li><font color="#0099FF">Ligation of July 15 [X/P] digested Rλ(co)-Rzλ(co) insert into the [S/P] digested pSB1C3-Sλmut(ori) vector</font></li><li><font color="#FF9900"> Ligation of July 15 [X/P] digested Rλ(co)-Rzλ(co) insert into the [S/P] digested pSB1C3-Sλmut(co) vector</font></li></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_Sλ(co), pGOv4-Ribo_Rλ(co) and pGOv4-Ribo_Rzλ(co) into competent <i>E. coli</i> cells</li><li>Transformation of pGOv4-SλWT(co)-Rλ(co)-Rzλ(co) (λLysis(c_c_c))</li></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-Sλmut(ori)-Rλ(co)-Rzλ(co) (λLysis_Sm(o_c_c))</font></li> <li><font color="#FF9900"> Transformation of the pSB1C3-Sλmut(co)-Rλ(co)-Rzλ(co) (λLysis_Sm(c_c_c))</font></li></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 Sλ(ori), Rλ(ori) and Rzλ(ori) genes using the extracted genomic DNA as template</font></li></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_Sλ(co) and pGOv4-SλWT(co)-Rλ(co)-Rzλ(co) (λLysis (c_c_c)) into competent <i>E. coli</i> cells</font></li></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 Rλ(co)-Rzλ(co) with [X/P] & Gel purification</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] & Gel purification</font></li></td>
    </tr>
    <tr>
      <td>Thursday 23</td>
-     <td><li><font color="#FF00FF">Extraction of the plasmids pGOv4-Ribo_Sλ(co), pGOv4-Ribo_Rλ(co) and pGOv4-Ribo_Rzλ(co)</li><li>Restriction digestion of the extracted plasmids with [E/P]</li><li>Gel purification</font></li></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 Rλ(co)-Rzλ(co) insert into the [S/P] digested pSB1C3-Sλmut(co) vector</li><li>Transformation</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_Sλ(co) and Ribo_Sλ(co) inserts into [E/P] digested pSB1C3 respectively</li><li>Transformation</font></li><br><li><font color="#3333FF">Restriction digestion of the Sλ(ori), Rλ(ori) and Rzλ(ori) amplicons with [X/P]</font></li></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 RRz(co) plasmid</font></li></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_Sλ and Ribo_Rλ clones</font></li><br><li><font color="#3333FF">Ligation of the [X/P] digested Sλ(ori), Rλ(ori) and Rzλ(ori) inserts into [X/P] digested pSB1C3 respectively</li><li>Transformation</font></li></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 & Gel purification</li><li><font color="#FF00FF">Restriction digestion of λLysis(c_c_c) with [X/P] & 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>