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

(Lab Work)
 
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{{Team:Paris_Saclay/notebook_header}}
 
{{Team:Paris_Saclay/notebook_header}}
=Tuesday 1<sup>st</sup> July=
+
=Friday 1<sup>st</sup> July=
==Lab Work==
+
  
 
===Biobrick characterization===
 
===Biobrick characterization===
 
====Buffer preparations====
 
====Buffer preparations====
''by Charlene and Caroline''
+
''By Charlene and Caroline''
  
 +
The following buffers were prepared:
 
{| class="wikitable"
 
{| class="wikitable"
 
! Buffer Z (250mL)
 
! Buffer Z (250mL)
! Buffer Z (250mL)
+
! Buffer Luc (250mL)
 
! Buffer STOP (15mL)
 
! Buffer STOP (15mL)
 
|-
 
|-
| Na2HPO4 – 3,125g
+
|  
NaH2PO4.H2O – 1,38g
+
Na<sub>2</sub>HPO<sub>4</sub> – 3,125g
  KCl – 0,185g
+
 
  MgSO4 – 0,04g
+
NaH<sub>2</sub>PO<sub>4</sub>.H<sub>2</sub>O – 1,38g
  β-mercaptoethanol – 0,893mL
+
 
| Trisphosphate pH 7,8 1M – 6,25mL
+
KCl – 0,185g
  MgCl2 2M – 1mL
+
 
  DTT 1M -0,25mL
+
MgSO<sub>4</sub> – 0,04g
  EDTA 0,5M – 0,5mL
+
 
  BSA – 0,25g
+
β-mercaptoethanol – 0,893mL
  Glycerol 60% - 62,5mL
+
 
  Triton 10% (100x) – 25mL
+
|  
| Na2CO3 1M – 0,78g
+
Trisphosphate pH 7,8 1M – 6,25mL
 +
 
 +
MgCl<sub>2</sub> 2M – 1mL
 +
 
 +
DTT 1M -0,25mL
 +
 
 +
EDTA 0,5M – 0,5mL
 +
 
 +
BSA – 0,25g
 +
 
 +
Glycerol 60% - 62,5mL
 +
 
 +
Triton 10% (100x) – 25mL
 +
|  
 +
Na<sub>2</sub>CO<sub>3</sub> 1M – 0,78g
 
|}
 
|}
 +
 +
===Visualization===
 +
====Migration of plasmids containing gBlocks digestion====
 +
''By Léa and Alice''
 +
 +
Digestion products from 30/06/16 were migrated on a gel (0.8% agarose) for 30 min at 100 V.
 +
[[File:T--Paris_Saclay--160701_visualization_gBlocks_échelle.jpg|400px|thumb|right|pPS16_002, pPS16_003 and pPS16_005 Migration]]
 +
[[File:T--Paris_Saclay--160701_visualization_gBlocks2_échelle.jpg#file|400px|thumb|right|pPS16_007, pPS16_008 and pPS16_009 Migration]]
 +
 +
{| class="wikitable"
 +
|-
 +
! Component
 +
! Volume (µL)
 +
|-
 +
| Digested DNA
 +
| 10
 +
|-
 +
| Loading buffer
 +
| 2
 +
|}
 +
 +
 +
For strains containing [[Team:Paris_Saclay/Notebook/June/28#pPS16_007|pPS16_007]] plasmid, according to the digestion products, 2 clones had the plasmid with the good insert. Indeed we could observe on the gel the two expected fragments of 2.7kb and 0.7kb. The same was observed for strains containing gBlocks 4.2 and GFP 1-9, respectively 3 and 2 clones were good. However, DNA concentration was too low.
 +
For others strains, migration products did not get consistent results, probably due to errors during the extraction.
 +
{{Team:Paris_Saclay/notebook_footer}}

Latest revision as of 13:56, 9 October 2016

Friday 1st July

Biobrick characterization

Buffer preparations

By Charlene and Caroline

The following buffers were prepared:

Buffer Z (250mL) Buffer Luc (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

By Léa and Alice

Digestion products from 30/06/16 were migrated on a gel (0.8% agarose) for 30 min at 100 V.

pPS16_002, pPS16_003 and pPS16_005 Migration
pPS16_007, pPS16_008 and pPS16_009 Migration
Component Volume (µL)
Digested DNA 10
Loading buffer 2


For strains containing pPS16_007 plasmid, according to the digestion products, 2 clones had the plasmid with the good insert. Indeed we could observe on the gel the two expected fragments of 2.7kb and 0.7kb. The same was observed for strains containing gBlocks 4.2 and GFP 1-9, respectively 3 and 2 clones were good. However, DNA concentration was too low. For others strains, migration products did not get consistent results, probably due to errors during the extraction.