Haynes Lab:Notebook/Short Projects/2015/02/23: Difference between revisions

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Line 33: Line 33:
| 10 uM fwd primer || 1.0 || 1.0 || 1.0  
| 10 uM fwd primer || 1.0 || 1.0 || 1.0  
|-
|-
| 10 uM rev primer || 1.0 || 1.0 || 1.0 || 1.0
| 10 uM rev primer || 1.0 || 1.0 || 1.0  
|-
|-
| 2x GoTaq green || 12.5 || 12.5 || 12.5 || 12.5  
| 2x GoTaq green || 12.5 || 12.5 || 12.5
|-
|-
| dH<sub>2</sub>O || 9.5 || 9.5 || 9.5
| dH<sub>2</sub>O || 9.5 || 9.5 || 9.5

Revision as of 11:24, 23 February 2015

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02/23/15 - Alexander Ellingson

PCR-verify activation domain primers
Try different cDNA libraries for the #name# fragment to account for any mutations or deletions

  1. U2OS C002, 1:1000 cDNA dilution; #name# = #size#: #f-primer#/ #r-primer#
  2. SKNSH C001, 1:1000 cDNA dilution; same
  3. K562 C001, 1:1000 cDNA dilution; same


Reagent Rxn1 Rxn2 Rxn3 Expected:
1. U2OS #name# = #size#
2. SKNSH #name# = #size#
2. K562 #name# = #size# 		Hover name
10 μL/lane, 1% agarose; Ladder
Template 1.0 1.0 1.0
10 uM fwd primer 1.0 1.0 1.0
10 uM rev primer 1.0 1.0 1.0
2x GoTaq green 12.5 12.5 12.5
dH2O 9.5 9.5 9.5
  25.0 25.0 25.0

Program: GOTAQ35cyc

  • 95°C, 3 min
  • 35x[95°C, 1 min; 57°C, 1 min; 72°C, 3 min]
  • 72°C, 3 min
  • 4°C ∞




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