Difference between revisions of "Team:Paris Saclay/Notebook/August/22"

(Created page with "{{Team:Paris_Saclay/notebook_header}} = Monday 22<sup>th</sup> August= ==Lab work== ===Visualization=== ====Low fidelity Dreamn Taq PCR of pPS16_016==== ''By Mathilde'' Clone...")
 
(Monday 22th August)
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* 1,5µL of Green Buffer
 
* 1,5µL of Green Buffer
 
* 1µL of dNTPs
 
* 1µL of dNTPs
* 1µL of each primers 1151 and 1151
+
* 1µL of each primers IPS83 and IPS84
 
* 0,13µL of Dream Taq Polymerase
 
* 0,13µL of Dream Taq Polymerase
 
* 19,37 µL of H20  
 
* 19,37 µL of H20  
  
Colonies were put into sterile water, and were put trhough
+
Colonies were put into sterile water, put through 5 minutes of denaturation before the addition of the mix.
 +
Annealing temperature was 57°c.
 +
After amplification, 1µL of each PCR product and 10µL of the DNA purple ladder were palced in wells and migrated at 100V for 30min.
 +
 
 +
====Purification on gel of gBlocks 3 and 4====
 +
''Terrence and Mathilde''
 +
 
 +
After amplification, 50µL of each PCR product diluted 6 tumes with 10 µL of the gel loadding dye, and 10µL of the DNA purple ladder were placed in wells and migrated at 100V for 30min.
  
  
  
 
{{Team:Paris_Saclay/notebook_footer}}
 
{{Team:Paris_Saclay/notebook_footer}}

Revision as of 13:18, 22 August 2016

Monday 22th August

Lab work

Visualization

Low fidelity Dreamn Taq PCR of pPS16_016

By Mathilde

Clones 7,8,9 and 10 of pPS16_016 were amplified following the protocol :

  • 1,5µL of Green Buffer
  • 1µL of dNTPs
  • 1µL of each primers IPS83 and IPS84
  • 0,13µL of Dream Taq Polymerase
  • 19,37 µL of H20

Colonies were put into sterile water, put through 5 minutes of denaturation before the addition of the mix. Annealing temperature was 57°c. After amplification, 1µL of each PCR product and 10µL of the DNA purple ladder were palced in wells and migrated at 100V for 30min.

Purification on gel of gBlocks 3 and 4

Terrence and Mathilde

After amplification, 50µL of each PCR product diluted 6 tumes with 10 µL of the gel loadding dye, and 10µL of the DNA purple ladder were placed in wells and migrated at 100V for 30min.