====Transformation of bacteria with [[Team:Paris_Saclay/Notebook/July/4#pPS16_004|pPS16_004]] and [[Team:Paris_Saclay/Notebook/June/28#pPS16_007|pPS16_007]] ====
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''By Alice and Terrence''
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''By Alice, Terrence and Mathilde''
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<!-- Rédigé par Terrence -->
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PCR made on bacteria transformed with pPS16_004 and pPS16_007 did not give expected results. We supposed that bacteria used for these PCRs did not still have the plasmid of interest. That is why we performed another transformation of DH5α competent cells with pPS16_004 and pPS16_007. We transformed cells following [[Team:Paris_Saclay/Experiments#Heat_shock_competent_cells|this protocol]]. We add 5μl of sterile H<SUB>2</SUB>O to the ligation product rest and used the whole for the transformation.
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As the result of PCRs on clone 2.2 and 4.1 didn't gave us expected results. We supposed that bacteria used for the PCR didn't integrate the plasmids. So we made another transformation of Dh5α competent cells.
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Finally, we spread cells on Petri dishes (LB + Amp 50μg/mL + xGal 0.25μL/mL + IPTG 0.1 μL/mL) in duplicate .
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We made a dilution with the rest of the ligation product with 5μl of sterile H<SUB>2</SUB>O and we follow the [[Team:Paris_Saclay/Experiments#Heat_shock_competent_cells|protocol of heat shock competent cells transformation]].
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Finally, we spread cells on Petri dishes (LB + Amp 50μg/mL + xGal 0.25μL/mL + IPTG0 0.1 μL/mL) in duplicate .
