Team:Freiburg/Notebook/1 September

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green light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME

blue light receptor

Picking clones

Investigators: Sandra

There were several clones on the cm plates. Clones were picked and incubated in LB cm medium at 37°C.

Gibson primer design

Investigators: Sophie, Sandra, Ruediger
we designed some new gibson primers for the assembly of NOT and LOV-tap in a vector because we are not sure if we can achieve this by 3A assemblies.

red light receptor

NAME OF YOUR EXPERIMENT

Investigators:NAME

Lysis cassette

3A Assembly of Quickchange-modified K098995 + modified Lysis genes K124017

Investigators:Theo
The digestion of the parts:

• S66 = modified Lysis genes K124017 in pSB1A2
• GFP = E0040 in pSB1A2
• S48 = Quickchange-modified K098995 in pSB1A2
• S39 = Promotor (J23104) + RBS (B0034) in pSB1C3

was run on the gel to see if the inserts are there.

There is no band for S66 so we sent it for sequencing

Precipitator

Ligation

Procedure

PCR tube:

total volume 20 μl

1. add H₂O (17 μl -X-Y-Z)
2. add 2 μl Ligase Buffer 10x
3. add Insert 1, Insert 2(when proceeding from 3A digestion use 2 μl of each)
4. add Vector (20ng needed. When proceeding from 3A digestion use 2 μl)
5. Add 1 μl T4-DNA Ligase
6. Incubate 10-30 min at room temperature
7. heat for 20 minutes at 80°C
8. store at -20°C or directly proceed to transformation

Documentation:

Why are you doing this experiment? Where are your parts stored? Name the parts for ligation etc.

Ligation

Procedure

PCR tube:

total volume 20 μl

1. add H₂O (17 μl -X-Y-Z)
2. add 2 μl Ligase Buffer 10x
3. add Insert 1, Insert 2(when proceeding from 3A digestion use 2 μl of each)
4. add Vector (20ng needed. When proceeding from 3A digestion use 2 μl)
5. Add 1 μl T4-DNA Ligase
6. Incubate 10-30 min at room temperature
7. heat for 20 minutes at 80°C
8. store at -20°C or directly proceed to transformation

Documentation:

Why are you doing this experiment? Where are your parts stored? Name the parts for ligation etc.