Contents |
Investigators:Jakob, Julia
Name:
Jakob | Date:
02.08.2011 |
Continue from Experiment 02.08.2011
Miniprep | |
Project Name: green light receptor, testdigest of CcaR with BamHI |
For one reaction you need: For Mastermix: Number of samples+2extra
4μl | H2O | 20 | |
1μl | Buffer, NEB4 | 5 | |
1μl | BSA (10x) | 5 | |
0,5 μl | Enzym 1 | 1 | BamHI |
0,5 μl | Enzym 2 | - | |
3 μl | DNA | 3 |
10 μl total volume
4μl | H2O | 20 | |
1μl | Buffer, NEB4 | 5 | |
1μl | BSA (10x) | 5 | |
0,5 μl | Enzym 1 | 1 | EcoRI |
0,5 μl | Enzym 2 | 1 | PstI |
3 μl | DNA | 3 |
10 μl total volume
Give 3 μl of DNA in an eppi and add 7μl of the mastermix.
Incubate for about 1h at 37°C.
Add 1 μl Loading dye buffer and load the gel.
Take a picture of the gel, print picture and label the lanes!
CcaR digested with BamH1 are negativ
Investigator:Jakob
PCR
Name:
Jakob | Date:
03.08.2011 |
Continue from Experiment: | |
Project Name:
Quickchange CcaR |
PCR-Mixture for one Reaction:
For a 50 µl reaction use
32,5µl | H20 | |
10µl | 5x Phusion Buffer | |
2.5µl | Primer fw | Primer: P12 |
2.5µl | Primer dw | Primer: P13 |
1µl | dNTPs | |
1µl | DNA-Template | DNA: CcaR |
0.5 µl | Phusion (add in the end) |
Investigators:Julia
transformation worked!
Investigators:NAME
Investigator:Jakob Picked some colonies for grow over night
Investigators:NAME
Investigators: NAME