Team:IONIS Paris/Notebook/18-08-15
Assembly of pDawn
Gibson assembly, NEB kit
Fragments | Concentration (ng.µL-1) | Lenght | Molecular weight (g.mol-1) | Molarity (mol.L-1) | 1/ratio | Dilution factor | Molarity after dilution | DNA Volume | Volume Water | Volume total |
---|---|---|---|---|---|---|---|---|---|---|
pSB1C3 | |
0.000000063713 | 0.000000031857 | |||||||
pDawnI | |
0.000000067280 | 0.000000063713 | |||||||
pDawnII | |
0.000000094981 | 0.000000063713 | |||||||
pDawnIII | |
0.000000112511 | 0.000000063713 |
DNA mix: 1µL of each diluted DNA solution or the indicated volume if no dilution is required (pDawnI) + 6,1 µL of MQ Water
Add 10 µL of master mix
50°C,15min
E.coli transformation: 1 aliquot with 5 µL of assembly product, 1 aliquot with 5 assembly product diluted by a factor 4.
18 August 15