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| | {{Team:Paris_Saclay/notebook_header}} | | {{Team:Paris_Saclay/notebook_header}} |
| − | =Thursday 7<sup>th</sup> July=
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| − | ==Lab work==
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| − | ===Visualization===
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| − | ====PCR products migation====
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| − | ''By Caroline and Léa''
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| − | gBlocks screened the [[Team:Paris_Saclay/Notebook/July/6#Visualization|06/07/2016]] were put on migration.
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| − | The usual [[Team:Paris_Saclay/Experiments#DNA_electrophoresis_on_agarose_gel|protocol]] was followed.
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| − | 20µL of each PCR product was added to 4µL of purple loading dye 6X.
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| − | PHOTO GEL pas dans le cahier !
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| − | ====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.
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| − | ====Culture of [[Team:Paris_Saclay/Notebook/July/6#Visualization|transformed]] pPS16_004, pPS16_007 and the pPS16_007 clone 6====
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| − | ''By Laetitia and Charlène''
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| − | Because of the absence of blue colony after culture, xGal and IPTG efficency was tested.
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| − | A petri dishe was splitted in four parts :
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| − | * new xGal, new IPTG
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| − | * new xGal, former IPTG
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| − | * former xGal, new IPTG
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| − | * 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===
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| − | ====BL21 pre-culture====
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| − | ''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====
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| − | ''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-).
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| − | For SP Cas9 (DS-SPcasN-) we used primers :
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| − | * IPS 134 + IPS 136 = SP1
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| − | * IPS 137 + IPS 135 = SP2
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| − | The PCR was conducted following the usual [[Team:Paris_Saclay/Experiments#Polymerase_chain_reaction|protocol]]
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| | {{Team:Paris_Saclay/notebook_footer}} | | {{Team:Paris_Saclay/notebook_footer}} |
