Difference between revisions of "Team:Paris Saclay/Notebook/August/11"
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= Thursday 11<sup>th</sup> August= | = Thursday 11<sup>th</sup> August= | ||
| − | + | ||
===Visualization=== | ===Visualization=== | ||
| − | ====Transformation of DH5a cells with | + | ====Transformation of DH5a cells with pPS16_020==== |
''By Léa'' | ''By Léa'' | ||
| − | Dh5a cells were transformed with | + | Dh5a cells were transformed with pPS16_020 (GFP 1-9 ligated with pSB1C3 digested), or the linearized pSB1C3 (control), or not transformed (control) using the usual [[Team:Paris_Saclay/Experiments#heat-shocktransformation|protocol]]. |
====[[Team:Paris_Saclay/Experiments#pPS16_010|pPS16_010]] extraction==== | ====[[Team:Paris_Saclay/Experiments#pPS16_010|pPS16_010]] extraction==== | ||
| Line 108: | Line 108: | ||
''By Laetitia'' | ''By Laetitia'' | ||
| − | Phusion PCR was performed on ligation product 2 ( | + | Phusion PCR was performed on the ligation product 2 (gBlock 2.1 ligated with gBblock 2.2) and the ligation product 3 (gBlock 3.1 ligated with gBlock 3.2) made on 09/08/16. |
It was done following [[Team:Paris_Saclay/Experiments#phusionPCR|this protocol]]. | It was done following [[Team:Paris_Saclay/Experiments#phusionPCR|this protocol]]. | ||
For the PCR on ligation 2 the primers used were IPS 123 and IPS 84. | For the PCR on ligation 2 the primers used were IPS 123 and IPS 84. | ||
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Gel1 | Gel1 | ||
| − | ====DreamTaq PCR of transformed cell with pPS16_002,008,010, 011, 012, 013, 014 and | + | ====DreamTaq PCR of transformed cell with pPS16_002,008,010, 011, 012, 013, 014 and puC19==== |
''By Léa and Laetitia'' | ''By Léa and Laetitia'' | ||
PCR DreamTaq was performed on 6 clones of each transformed cells from 10/08/16. | PCR DreamTaq was performed on 6 clones of each transformed cells from 10/08/16. | ||
| − | Each clone was | + | Each clone was spread and put in liquid culture with LB and Ampicillin. |
No PCR products were observed on the gel. | No PCR products were observed on the gel. | ||
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''By Alice'' | ''By Alice'' | ||
| − | + | pSB1C3 was amplified with Phusion polymerase following [[Team:Paris_Saclay/Experiments#phusionPCR|this protocol]]. Two primers ([[Team:Paris_Saclay/Experiments#primers|iPS41 and iPS42]]) were used. Annealing temperature was 70.9°C. After amplification, 5 µL of each PCR products with loading dye and 10 µL of DNA ladder were placed on gel and migrated at 100v during 30 min. | |
PCR products expected: | PCR products expected: | ||
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''By Caroline'' | ''By Caroline'' | ||
| − | We made the PCR again as the previous result | + | We made the PCR again as the previous result was not satisfying. |
Ligation product were cultivate overnight. | Ligation product were cultivate overnight. | ||
[[File:T--Paris_Saclay--extraction_20160811_pcr_ligation2-3.jpg|500px|thumb|right|Result of the ligation 2-3 PCR]] | [[File:T--Paris_Saclay--extraction_20160811_pcr_ligation2-3.jpg|500px|thumb|right|Result of the ligation 2-3 PCR]] | ||
===Bringing DNA closer === | ===Bringing DNA closer === | ||
| − | + | ==== Extraction of DS-SPcasN- and DS-TDcasN- ==== | |
| − | ==== Extraction of DS-SPcasN- | + | |
''By Terrence'' | ''By Terrence'' | ||
Latest revision as of 16:37, 9 October 2016
