====High fidelity PCR on bacteria transformed with [[Team:Paris_Saclay/Notebook/June/28#pPS16_005|pPS16_005]] ====
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''By ''
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''By Alice''
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High fidelity PCR on bacteria transformed with pPS16_005 did not get PCR products. We supposed that there was a problem with [[Team:Paris_Saclay/Experiments#primers|iPS83 primer]] dilution. That is why we performed again this PCR after diluting again theses primers. We followed [[Team:Paris_Saclay/Experiments#Q5PCR|the same protocol]] as previously. [[Team:Paris_Saclay/Experiments#primers|iPS83 and iPS126]] primers were used. Annealing temperature was 72°C, and initial step was prolonged to 7 min. After amplification, 1 µL of loading dye was added to 5µL of each PCR products. Then 5 µL of this mix were put into a well of the gel. A last well contained 10µL of DNA ladder. PCR products were migrated 25min at 100V.
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PCR products expected were :
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''By ''
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{| class="wikitable"
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|-
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!Plasmids
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!Band size (bp)
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|-
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|pPS16_005
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|964
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|}
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We did not observed PCR products. This is maybe due to a iPS83 primer default. New primers iPS83 will be ordered to perform again this PCR.
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====Culture of bacteria transformed with pPS16_002 and pPS16_004====
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''By Alice''
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After transformation made on the [[Team:Paris_Saclay/Notebook/July/19#transformation__4.1_2.2_1.2|19/07/16]], bacteria were spread on petri dishes without working XGal and IPTG, explaining that we get white colonies only. These colonies were spread again on petri dishes with LB, Ampicilin (50µg/mL), XGal (0.25µL/mL) and IPTG (0.1µL/mL).
