(→Gibson of cleaned up PCR products fragment 1 and fragment 2 in pSB1C3 treated by DpnI)
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[[Team:Paris_Saclay/Experiments#primers|Primers]] used were:
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[[Team:Paris_Saclay/Experiments#primers|Primers]] used were:
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|Primers
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|iPS122 and Ips140
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|iPS122 and iPS140
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|iPS123 and Ips84
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|iPS123 and iPS84
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|Tm
|Tm
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[[File:T--Paris_saclay--0509.extension.png|400px|thumb|center|Result of migration ]]
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The PCR product of ligation is not in high concentration (mostly with phusion polymerase)
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Following Gibson assembly was done by both fragments.
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====Gibson of cleaned up PCR products fragment 1 and fragment 2 in pSB1C3 treated by DpnI====
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''By Manhaz ''
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Gibson was performed with cleaned up PCR products fragment1 x fragment 2 x pSB1C3 treated by DpnI (plasmid) with the following protocol:
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For each 20μl of reaction, mix the following reagents :
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* the 1-2 Gibson assembly product tube : 1,12µL of purified segment 1 and 0,56µL of purified segment 2 + 0,85µL of digested pSB1C3 + 7,47 µL of sterile water + 10µL of NEB Builder Hifi DNA Assembler Master Mix
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* the control tube : 0,85µL of digested pSB1C3 + 17,47 µL of sterile water
