"

From 2011.igem.org

Contents 1 Meeting 1.1 green light receptor 1.2 blue light receptor 1.3 red light receptor 1.4 Lysis cassette 1.5 Quick change 2 Precipitator

Meeting

attendants: Tobi, Theo, Julia, Rüdiger, Jakob, Sandra

green light receptor

already done:

To-do:

blue light receptor

already done:

1. Transformation of Lov-Tap in cells.

To-do:

1. Cloning of promotor (medium) in Lov-Tap part and perform a transformation. Plating out of the cells on trypthophan-free medium.

red light receptor

already done:

To-do:

Lysis cassette

Quick change

Investigators: Theo

already done:

1. repressor part (BBa_K098995) has no bases between RBS (B0034) and start codon (K098997), resulting in no translation.
   • quickchange of the repressor part to insert 6bp between RBS and start codon

To-do:

1. DpnI digestion to digest the DNA template (methylated DNA) of the PCR to have only the new synthesized DNA strand.
2. After digestion with DpnI, transformation of cells with the corrected part (BBa_K08995)

Precipitator

PCR

Name: Ruediger	Date: 18.07.2011
Continue from Experiment (Date)PCR 1507 (Name) Ruediger
Project Name: GFP Pbd

PCR-Mixture for one Reaction:

For a 50 µl reaction use

32,5µl	H20
10µl	5x Phusion Buffer
2.5µl	Primer fw
2.5µl	Primer dw
1µl	dNTPs
1µl	DNA-Template
0.5 µl	Phusion (add in the end)

What program do you use?

10x (95C-41/52C-72C) + 25x ((95C-60C-72C)

How did you label the PCR-Product, where is it stored and what do you do next?

Reactions:

lane 3

P28+P18+M14.1

lane 4

P28+P19+M14.1

lane 5

P28+P20+M14.1

Results:

did not work well.

Strange band size in lane 3. 4 and 5 did not work.