(→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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The PCR product of ligation is not in high concentration (mostly with phusion polymerase)
The PCR product of ligation is not in high concentration (mostly with phusion polymerase)
Following Gibson assembly was done by both fragments.
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
