Template:IONIS Paris/Notebook/18-08-15

Assembly of pDawn

Gibson assembly, NEB kit

Quantification of DNA fragments by Image J:

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	91.11	2 200	1 430 000	0.000000063713	1.77	1.00	0.000000031857	1.00	1.00	2.00
pDawnI	45.70	1045.00	679 250	0.000000067280	1.67	1.06	0.000000063713	0.95	0.00	0.95
pDawnII	54.45	882	573 300	0.000000094981	1.18	1.49	0.000000063713	2.00	0.98	2.98
pDawnIII	119.57	1635	1 062 275	0.000000112511	1.00	1.77	0.000000063713	1.00	0.90	1.90

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 August15