Difference between revisions of "Team:Paris Saclay/Notebook/August/29"

(Created page with "PCR sur colonie ====PCR of the plasmid extraction from clonies 4,5,9,12 containing FRB sequence in pJET plasmid==== ''Mahnaz'' * 2.5 µL DreamTaq Buffer * 0.5 µL of dNTPs (...")
 
(→‎PCR of the plasmid extraction from clonies 4,5,9,12 containing FRB sequence in pJET plasmid)
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====PCR of the plasmid extraction from clonies 4,5,9,12 containing FRB sequence in pJET plasmid====
 
====PCR of the plasmid extraction from clonies 4,5,9,12 containing FRB sequence in pJET plasmid====
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''Mahnaz''
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''By Mahnaz''
  
 
* 2.5 µL DreamTaq Buffer
 
* 2.5 µL DreamTaq Buffer
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|30 sec
 
|30 sec
 
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|-
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|Tm
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|52
 
|30 sec
 
|30 sec
 
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|52°C
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|72°C
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|t
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|1 min
 
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|Final Extension
 
|Final Extension
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|Hold
 
|Hold
 
|4°C
 
|4°C
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|$\infinity\$
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|\infinity\
 
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|Primers
 
|Primers
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|iPS83 and iPS84
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|pJET R and F
 
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|Tm
 
|Tm
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|63.6°C
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|°C
 
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|t
 
|t

Revision as of 08:13, 16 September 2016

PCR sur colonie

PCR of the plasmid extraction from clonies 4,5,9,12 containing FRB sequence in pJET plasmid

By Mahnaz

  • 2.5 µL DreamTaq Buffer
  • 0.5 µL of dNTPs (10mM)
  • 1 µL of each primer (10µM) primer pJET R and F
  • 0.13 μl of DreamTaq Pol
  • 20 µl H2O

PCR was performed as follow:

Step Temperature Time
Initial denaturation 95°C 3 min
30 cycles 95°C 30 sec
52 30 sec
72°C 1 min
Final Extension 72°C 7min
Hold 4°C \infinity\ 
Primers used were:
Matrix Clones containing dCas9 NM - GFP 10 in pSB1C3
Primers pJET R and F
Tm °C
t 1 min 30

After amplification, 3 µL of each PCR products and 5 µL of DNA ladder were placed in wells and migrated at 100V during 30 min.

PCR products expected were :

PCR products Expected band size (bp)
dCas9 NM - GFP 10 - pSB1C3 3688
Result of the migration

GEL GEL GEL