Difference between revisions of "Team:Paris Saclay/Notebook/August/5"

(→‎Q5 PCR on DH5alp|pPS16_003, DH5alp|pPS16_004, DH5alp|pPS16_006 and DH5alp|pPS16_007)
(→‎Q5 PCR on pPS16_003, pPS16_004, pPS16_006 and pPS16_007)
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The PCR was carried out following the usual [[Team:Paris_Saclay/Experiments#Q5PCR|protocol]] adapted to 50µL and with a TM at 70°C. The specific [[Team:Paris_Saclay/Experiments#primers|primers]] for each parts were using. The products were put to migrated on a 0.8%agarose gel with BET. Only, PCR form pPS16_003 and pPS16_004 showed a positive result.
 
The PCR was carried out following the usual [[Team:Paris_Saclay/Experiments#Q5PCR|protocol]] adapted to 50µL and with a TM at 70°C. The specific [[Team:Paris_Saclay/Experiments#primers|primers]] for each parts were using. The products were put to migrated on a 0.8%agarose gel with BET. Only, PCR form pPS16_003 and pPS16_004 showed a positive result.
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==== PCR Clean-up with the NucleoSpin kit ====
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''By Caroline''
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The purification was carried out on DH5alp|pPS16_003, DH5alp|pPS16_004 and DH5alp|pPS16_007 following the usual [[Team:Paris_Saclay/Experiments#Purification|protocol]]. But, I forgot to diluted the Buffet NT3 with 100mL ethanol befor I began the purification and so any result was obtained on the 0.8% agarose gel.

Revision as of 14:03, 5 August 2016

Friday 5st August

Lab work

Visualization

DNA Extraction of DS-TDcasN- and DS-SPcasN-

By Laetitia


The extraction was performed using the kit. We followed this protocol.

The initial culture was at 15 mL so we increased (*4) the volumes until the precipitation.

Hence we used:

-1 ml of resuspension buffer

-1 ml lysis buffer

-1 ml precipitation buffer

The colum was used several times in order to recover the maximum of DNA

DNA Extraction of DS-NMcasN- and pPS_001

By Mathilde

The extraction was performed on clone 11 for NM and clone 9 for 1.1 using the kit. We followed this protocol.

Q5 PCR on pPS16_003, pPS16_004, pPS16_006 and pPS16_007

By Caroline

The PCR was carried out following the usual protocol adapted to 50µL and with a TM at 70°C. The specific primers for each parts were using. The products were put to migrated on a 0.8%agarose gel with BET. Only, PCR form pPS16_003 and pPS16_004 showed a positive result.

PCR Clean-up with the NucleoSpin kit

By Caroline

The purification was carried out on DH5alp|pPS16_003, DH5alp|pPS16_004 and DH5alp|pPS16_007 following the usual protocol. But, I forgot to diluted the Buffet NT3 with 100mL ethanol befor I began the purification and so any result was obtained on the 0.8% agarose gel.