Haynes Lab:Notebook/Engineering PC-TFs/2012/12/06: Difference between revisions

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Run PCR with preset GoTaq settings. Need to ask Dr. Haynes to be certain settings are correct for my reactions.
Run PCR with preset GoTaq settings. Need to ask Dr. Haynes to be certain settings are correct for my reactions.<br>
 
______________________
  *Digest the pSB1A2/3 plasmid (from Rene) with XbaI & SpeI to get rid of the insert
  *Digest the pSB1A2/3 plasmid (from Rene) with XbaI & SpeI to get rid of the insert


Revision as of 16:20, 6 December 2012

Engineering PC-TFs <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
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Summary

Dilute primers that came from IDT (10x dilution of concentration)

  • BL_Primer09 26.6 nm --> 266 μL dilution
  • BL_Primer10 25.7 nm --> 257 μL dilution
  • Thaw and dilute primers that came (x10 dilution)
  • 90 μL water
  • 10 μL primer stock

(10 μm in each tube for PCR reaction)
______________________________________

  • Assemblies

1. hPCD - Pflex - BL01
2. hPCD - BL01

# Template Primers
1 hPCD 9, 2
2 Plflex 3, 4
3 BL01 5,10
4 hPCD 9, 7
5 BL01 8,10
μL
Template .5 μL
Primer 1 (10 μm) 1.0 μL
Primer 2 (10 μm) 1.0 μL
2*GoTaq 12. 5
DH2O 10.0 μL

Run PCR with preset GoTaq settings. Need to ask Dr. Haynes to be certain settings are correct for my reactions.
______________________ 

*Digest the pSB1A2/3 plasmid (from Rene) with XbaI & SpeI to get rid of the insert
Reagent Volume  
DNA (plasmid) 20.0
10x buffer 3.0
XbaI 1.0
SpeI 1.0
dH2O 5.0
  30 μL --> 37°C/ ~10 min.

Vi helped me with running gel because I had class. Vi will put digest into gel and will get results.



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