Difference between revisions of "Team:Paris Saclay/Notebook/July/1"

Line 43: Line 43:
 
===Visualization===
 
===Visualization===
 
====Migration of plasmids containing gBlocks digestion====
 
====Migration of plasmids containing gBlocks digestion====
−
Products of the digestion made the 30/06/16 are migrated on a gel (0.8% agarose) during 30 min, 100 V.
+
Products of the digestion made the 30/06/16 were migrated on a gel (0.8% agarose) during 30 min, 100 V.
  
 
{| class="wikitable"
 
{| class="wikitable"
Line 56: Line 56:
 
| 2
 
| 2
 
|}
 
|}
 +
 +
For strains containing plasmids with gBlocks 4.1, according to the digestion products, 2 clones had the plasmid with the good insert. Indeed we could observe on the gel, the two fragments expected (2.7 and 0.7 Kb). All the same for strains containing gBlocks 4.2 and GFP 1-9, 3 and 2 clones respectively were good. However, DNA concentration is too low.
 +
For others strains, migration products did not get consistent results, probably due to errors during the extraction.

Revision as of 10:00, 7 July 2016

Tuesday 1st July

Lab Work

Biobrick characterization

Buffer preparations

by Charlene and Caroline

Buffer Z (250mL) Buffer Z (250mL) Buffer STOP (15mL)

Na2HPO4 – 3,125g

NaH2PO4.H2O – 1,38g

KCl – 0,185g

MgSO4 – 0,04g

β-mercaptoethanol – 0,893mL

Trisphosphate pH 7,8 1M – 6,25mL

MgCl2 2M – 1mL

DTT 1M -0,25mL

EDTA 0,5M – 0,5mL

BSA – 0,25g

Glycerol 60% - 62,5mL

Triton 10% (100x) – 25mL

Na2CO3 1M – 0,78g

Visualization

Migration of plasmids containing gBlocks digestion

Products of the digestion made the 30/06/16 were migrated on a gel (0.8% agarose) during 30 min, 100 V.

Component Volume (µL)
Digested DNA 10
Loading buffer 2

For strains containing plasmids with gBlocks 4.1, according to the digestion products, 2 clones had the plasmid with the good insert. Indeed we could observe on the gel, the two fragments expected (2.7 and 0.7 Kb). All the same for strains containing gBlocks 4.2 and GFP 1-9, 3 and 2 clones respectively were good. However, DNA concentration is too low. For others strains, migration products did not get consistent results, probably due to errors during the extraction.