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

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(Autocreate 2015/02/23 Entry for Haynes_Lab:Notebook/Short_Projects)
 
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==mm/dd/yy - First Last==
==02/23/15 - Alexander Ellingson==
* Insert content here...
 
<!-- Replace #name# with activation domain name.
 
'''PCR-verify fragment primers'''<br>
Try different cDNA libraries for the #name# fragment to account for any mutations or deletions
# U2OS C002, 1:1000 cDNA dilution; #name# = #size#: ATF2_f1/ ATF2_r1
# SKNSH C001, 1:1000 cDNA dilution; same
# K562 C001, 1:1000 cDNA dilution; same
 
 
{| {{table}} cellspacing="3" <!-- PCR rxn. table -->
|- valign="top"
| bgcolor=#cfcfcf | Reagent
| bgcolor=#cfcfcf | Rxn1
| bgcolor=#cfcfcf | Rxn2
| bgcolor=#cfcfcf | Rxn3
| bgcolor=#cfcfcf | Rxn4
| rowspan="7" | Expected:<br>1. KAH60 = 6637<br>2. ATF2 = 906<br>3. ATF2 = 906<br>4. ATF2 = 906
| rowspan="7" | [[Image:KAH012315_gel.jpg|270px|Hover name]]<br>10 μL/lane, 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
|-
| Template || 0.2 || 1.0 || 1.0 || 1.0
|-
| 10 uM fwd primer || 1.0 || 1.0 || 1.0 || 1.0
|-
| 10 uM rev primer || 1.0 || 1.0 || 1.0 || 1.0
|-
| 2x GoTaq green || 12.5 || 12.5 || 12.5 || 12.5
|-
| dH<sub>2</sub>O || 10.3 || 9.5 || 9.5 || 9.5
|-
| &nbsp; || 25.0 || 25.0 || 25.0 || 25.0
|}
 
Program: GOTAQ35cyc
* 95°C, 3 min
* 30x[95°C, 1 min; 57°C, 1 min; 72°C, 3 min]
* 72°C, 3 min
* 4°C ∞
 
 




Revision as of 11:19, 23 February 2015

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02/23/15 - Alexander Ellingson

Reagent Rxn1 Rxn2 Rxn3 Rxn4 Expected:
1. KAH60 = 6637
2. ATF2 = 906
3. ATF2 = 906
4. ATF2 = 906 		Hover name
10 μL/lane, 1% agarose; Ladder
Template 0.2 1.0 1.0 1.0
10 uM fwd primer 1.0 1.0 1.0 1.0
10 uM rev primer 1.0 1.0 1.0 1.0
2x GoTaq green 12.5 12.5 12.5 12.5
dH2O 10.3 9.5 9.5 9.5
  25.0 25.0 25.0 25.0

Program: GOTAQ35cyc

  • 95°C, 3 min
  • 30x[95°C, 1 min; 57°C, 1 min; 72°C, 3 min]
  • 72°C, 3 min
  • 4°C ∞



|}


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