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

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