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Team:Freiburg/Notebook/6 September
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==<span style="color:orange;">Lysis cassette</span>== | ==<span style="color:orange;">Lysis cassette</span>== | ||
| - | === | + | ===Troubleshooting of the modified Lysis genes K124017=== |
| - | '''Investigators: | + | '''Investigators:Theo''' |
| + | <br> | ||
| + | Minipreps of the first inoculations were applied on a 1% agarose gel without digesting them to see if they have an insert of the modified Lysis genes K124017. Nr1, 5 and 7 were sent for sequencing since the size seems to be about right. | ||
| + | *Note: S4 is the B0034 RBS (ribosome binding site), S15 is the non-modified K124017 Lysis Cassette without RBS | ||
| + | <br> | ||
| + | |||
| + | [[File:Freiburg11_9_7_2011_Gel_von_S4plusS15_ohne_Verdau.Jpg|700px]] | ||
| + | |||
| + | <br> | ||
==<span style="color:grey;">Precipitator</span>== | ==<span style="color:grey;">Precipitator</span>== | ||
Revision as of 18:57, 6 September 2011
Contact 
Contents |
Commons
Miniprep
Investigators: Sandra
Yesterday LB was inoculated with strains (S14, S39-S44 etc.). Miniprep of strains.
Digestion: 2A-assembly
Investigators: Sandra
Digestion of:
- S14 (GFP)
with XbaI and PstI
- S39 (PR1)
- S40 (PR2)
- S41 (PR3)
- S42 (PR4)
- S43 (PR5)
- S44 (PR6)
with SpeI and PstI
incubation for 6 hours at 37°C.
green light receptor
NAME OF YOUR EXPERIMENT
Investigators:NAME
blue light receptor
Trafo
Investigators: Sandra
Trafo of ligated parts of the 2A-assembly.
Sequence analysis
sequence analysis of our lovtap-Notgate was neagtive:-( So we´ll try the gibson assembly.
PCR
Investigators: Sophie PCR
| Name: Sophie
| Date: 06.09.11 |
| Continue from Experiment (Date)
(Name) | |
| Project Name: Gibson of ♥ and NOT | |
PCR-Mixture for one Reaction:
For a 50 µl reaction use
| 32,5µl | H20 | Name |
| 10µl | 5x Phusion Buffer | of Primer |
| 2.5µl | Primer fw | ♥: P97
NOT: P99 |
| 2.5µl | Primer dw | ♥: P98
NOT: P100 |
| 1µl | dNTPs | of Template DNA |
| 1µl | DNA-Template | NoT, M45 (BBa_Q0400)
original DNA from Edinburgh (BBa_K322999) |
| 0.5 µl | Phusion (add in the end) |
Digestion with Dpn I and purification with PCR purification kit What program do you use?
First 20 cycles touchdown 69°C -0.4/cycle
next 10 cycles touchdown 72°C -0.2 per cycle
To confirm the PCR-Product has the correct size, load 2 µl of the sample onto an agarose-gel.
How did you label the PCR-Product, where is it stored and what do you do next?
red light receptor
NAME OF YOUR EXPERIMENT
Investigators:NAME
Lysis cassette
Troubleshooting of the modified Lysis genes K124017
Investigators:Theo
Minipreps of the first inoculations were applied on a 1% agarose gel without digesting them to see if they have an insert of the modified Lysis genes K124017. Nr1, 5 and 7 were sent for sequencing since the size seems to be about right.
- Note: S4 is the B0034 RBS (ribosome binding site), S15 is the non-modified K124017 Lysis Cassette without RBS
Precipitator
NAME OF YOUR EXPERIMENT
Investigators: NAME
