Friday 7th August

Lab Work

Transformation

by Coralie

BBa_K1707032
BBa_K1707033
BBa_K1707026

As usual

Plasmid extraction

by Pauline

BBa_R0040 #1 and #2
BBa_K1707020 #1 and #2
BBa_K1707027 #1 and #2
BBa_K1707028 #1 and #2
Plasmid test #1 and #2

With Macherey-Nagel Extraction kit

Digestion verification

by Pauline

BBa_K1707020 #1 and #2
BBa_K1707027 #1 and #2
BBa_K1707028#1 and #2
Plasmid test #1 and #2

Mix:

2 µL plasmid
0,5 µL XbaI
0,5 µL PstI
1µL Buffer FastDigest 10x
6 µL H2O

Incubation 1h30, 37°C

SpeI Test

by Pauline

Mix:

2 µL BBa_K1707028 #1
0,5 µL SpeI
1µL Buffer N
6,5 µL H2O

Incubation 2h30, 37°C

Electrophoresis

by Pauline

Agarose gel 1%, 100V

Well 1: Test of SpeI, Wells 3-10: Verification by digestion with XbaI and PstI; from left to right: 1. Test SpeI, 2. DNA Ladder, 3. BBa_K1707030#1, 4. BBa_K1707030#2, 5. BBa_K1707020#1, 6. BBa_K1707020#2, 7. BBa_K1707027#1, 8. BBa_K1707027#2, 9. K1707028#1, 10. K1707028#2

We can conlude that:

SpeI is OK
BBa_K1707020 #1 isn't OK
BBa_K1707020 #2 is OK
BBa_K1707027 #1 and #2 are OK
BBa_K1707028 #1 and #2: we have to check them another time
Plasmid test #1 and #2 are OK but not totally digested

New culture

by Pauline

BBa_R0051 #1

with 5mL LB and 5 µL Ampicillin

Digestion Verification

by Pauline

Mix:

2 µL Plasmid
0,5 µL XbaI
0,5 µL PstI
6 µL H2O
1 µL Buffer FastDigest 10x

Low budget challenge

by Coralie

We can see a lot of small colony. We put it ON 37°C.

Member present:

Instructors: Claire
Students: Coralie and Pauline

Back to the calendar

Team:Paris Saclay/Notebook/August/7

Contents

Friday 7th August

Lab Work

Transformation

Plasmid extraction

Digestion verification

SpeI Test

Electrophoresis

New culture

Digestion Verification

Low budget challenge