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

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Digest the pSB1A2/3 plasmid (from Rene) with XbaI & SpeI to get rid of the insert
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{| class="wikitable" border="0" cellspacing="3" <!-- Digest rxn. table -->
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|-valign="top"
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| <u>Reagent</u> || <u>Volume</u> || &nbsp;
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| rowspan="9" |
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|-
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| DNA (plasmid) || 20.0
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| 10x buffer || 3.0
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|-
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| XbaI || 1.0
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|-
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| SpeI || 1.0
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|-
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| dH<sub>2</sub>O || 5.0
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|-
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| &nbsp; || 30 μL --> 37°C/ ~10 min.
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|}
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Vi helped me with running gel because I had class. Vi will put digest into gel and will get results.

Revision as of 19:17, 6 December 2012

Engineering PC-TFs 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 
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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