Difference between revisions of "Team:Paris Saclay/Notebook"
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The usual [[Team:Paris_Saclay/Experiments#DNA_electrophoresis_on_agarose_gel|protocol]] was followed. | The usual [[Team:Paris_Saclay/Experiments#DNA_electrophoresis_on_agarose_gel|protocol]] was followed. | ||
20µL of each PCR product was added to 4µL of purple loading dye 6X. | 20µL of each PCR product was added to 4µL of purple loading dye 6X. | ||
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PHOTO GEL pas dans le cahier ! | PHOTO GEL pas dans le cahier ! | ||
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====Cultures results==== | ====Cultures results==== | ||
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There was no blue colony observed on xGal/IPTG + ampicillin mediums for transformed [[Team:Paris_Saclay/Notebook/July/6#Visualization|transformed]] pPS16_004, pPS16_007 and the pPS16_007 clone 6 gBlocks. | There was no blue colony observed on xGal/IPTG + ampicillin mediums for transformed [[Team:Paris_Saclay/Notebook/July/6#Visualization|transformed]] pPS16_004, pPS16_007 and the pPS16_007 clone 6 gBlocks. | ||
| + | ====Culture of [[Team:Paris_Saclay/Notebook/July/6#Visualization|transformed]] pPS16_004, pPS16_007 and the pPS16_007 clone 6==== | ||
| + | ''By Laetitia and Charlène'' | ||
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| + | Because of the absence of blue colony after culture, xGal and IPTG efficency was tested. | ||
| + | A petri dishe was splitted in four parts : | ||
| + | * new xGal, new IPTG | ||
| + | * new xGal, former IPTG | ||
| + | * former xGal, new IPTG | ||
| + | * former xGal, former IPTG | ||
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| + | A blue colony from the control tube 2 of the [[Team:Paris_Saclay/Notebook/July/4#Visualization|04/07/2016]] was plated on each of the four part on medium LB + Ampicillin (100µg/mL) + xGal/IPTG (1/1000). | ||
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| + | 100µ of pPS16_004, pPS16_007 or the pPS16_007 clone 6 bacteries were plated again on LB + Ampicillin (100µg/mL) + xGal/IPTG (1/1000) medium. | ||
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| + | ===caracterization=== | ||
| + | ====BL21 pre-culture==== | ||
| + | ''By Laetitia and Charlène'' | ||
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| + | A BL21 colony was added to 4µL of LB and put in incubation overnight at 37°c, 200 rpm. | ||
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| + | ====Cas9 PCR==== | ||
| + | ''By Léa'' | ||
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| + | Forward primer Sequencing addgene Cas plasmids (IPS 134) and Reverse primer Sequencing addgene Cas plasmids (IPS 135) were used for NM Cas9(DS-NMcas), ST1 Cas9 (DS-ST1casN-) and Td Cas9 (DS-TDcasN-). | ||
| + | For SP Cas9 (DS-SPcasN-) we used primers : | ||
| + | * IPS 134 + IPS 136 = SP1 | ||
| + | * IPS 137 + IPS 135 = SP2 | ||
| + | The PCR was conducted following the usual [[Team:Paris_Saclay/Experiments#Polymerase_chain_reaction|protocol]] | ||
Revision as of 10:19, 19 July 2016
