Difference between revisions of "Team:Paris Saclay/Notebook/August/12"
(→Extraction of puc19) |
(→Lab work) |
||
| (18 intermediate revisions by 6 users not shown) | |||
| Line 2: | Line 2: | ||
= Friday 12<sup>th</sup> August= | = Friday 12<sup>th</sup> August= | ||
| − | + | ||
===Visualization=== | ===Visualization=== | ||
| − | ====PCR of | + | ====PCR of pPS16_020==== |
''By Léa'' | ''By Léa'' | ||
| Line 10: | Line 10: | ||
Primers: IPS83 ans IPS84 | Primers: IPS83 ans IPS84 | ||
| − | ====Extraction of | + | [[File:T--Paris_Saclay--120826_visualization_PCRGFP.jpg|400px|thumb|right|Electrophoresis of PCR products, using primers IPS 83 and IPS 84 on pPS16_016.]] |
| + | |||
| + | ====Extraction of pUC19==== | ||
''By Charlène'' | ''By Charlène'' | ||
| − | Clones 1 and 2 of | + | Clones 1 and 2 of pUC19 were extracted with the [[Team:Paris_Saclay/Experiments#InvitrogenPlasmidExtraction|Plasmid MiniPrep kit]]. |
| − | They were fast digested with EcoRI : 3µL of plasmid + 1µL of FD buffer + 0.5µL of FD EcoRI + 5.5µL of water, 15 minutes at 37°C. | + | |
| + | They were fast digested with EcoRI : 3µL of plasmid + 1µL of FD buffer + 0.5µL of FD EcoRI + 5.5µL of water, 15 minutes at 37°C. | ||
| + | |||
| + | [[File:T--Paris_Saclay--20160812_puc19_digestion.JPG|500px|thumb|right|pUC19 clones 1 and 2 digested by EcoRI]] | ||
| + | |||
| + | |||
| + | Just clone 1 is at the good size so we will have to use it and not clone 2. | ||
| + | |||
| + | ====Dreamtaq PCR on puc19, detection, FRB, FKBP, SgRNAst, SgRNAnm and gBlock 1.2==== | ||
| + | ''By Naiane, Mahnaz and Terrence'' | ||
| + | |||
| + | A Dreamtaq PCR was performed on puc19, detection, FRB, FKBP, SgRNAst, SgRNAnm and gBlock 1.2 using the usual [[Team:Paris_Saclay/Experiments#taqPCR|protocol]] with these volumes : | ||
| + | |||
| + | {| class="wikitable" | ||
| + | |- | ||
| + | !Components | ||
| + | !Volume | ||
| + | |- | ||
| + | |10X DreamTaq Green Buffer | ||
| + | |2.5µL | ||
| + | |- | ||
| + | |dNTP (10mM) | ||
| + | |1µL | ||
| + | |- | ||
| + | |Primers mix (10µM each) | ||
| + | |1µL | ||
| + | |- | ||
| + | |DreamTaq DNA polymerase | ||
| + | |0.25µL | ||
| + | |- | ||
| + | |Nuclease-free water | ||
| + | |up to 25µL | ||
| + | |- | ||
| + | !Total volume | ||
| + | !25µL | ||
| + | |} | ||
| + | We made 6 clones of each colonie except for Puc19 where we made 1 negative control. | ||
| + | |||
| + | Result of the PCR : there is no band at the expected size for all the screened clones so the transformations of the gblock do not seem to have worked. | ||
| + | |||
| + | [[File:T--Paris Saclay--1-2-FRB 15 08.jpeg|400px|thumb|right|1.2 and FRB]] | ||
| + | |||
| + | [[File:T--Paris_Saclay--détection-ST_15_08.JPG|400px|thumb|right|Detection and ST]] | ||
| + | |||
| + | |||
| + | [[File:T--Paris_Saclay--NM-FKBP_15_08.JPG|400px|thumb|right|puc19, NM and FKBP]] | ||
| + | |||
| + | |||
| + | [[File:T--Paris_Saclay--puc19-FKBP_15_08.JPG|400px|thumb|right|FKBP]] | ||
| + | |||
| + | |||
| + | |||
| + | |||
{{Team:Paris_Saclay/notebook_footer}} | {{Team:Paris_Saclay/notebook_footer}} | ||
Latest revision as of 16:39, 9 October 2016
