Haynes Lab:Notebook/Jan/2016/06/23: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
Line 14: Line 14:
| bgcolor=#cfcfcf | Mix* (x #)
| bgcolor=#cfcfcf | Mix* (x #)
| rowspan=7 | Expected:<br>1. 4-x, P1 = 1145 <br>2. 4-x, P3 = 1046 <br>3. 4-x, P6 = 1139 <br>4. 6-x, P1 = 1146 <br>5. 6-x, P3 = 1046 <br>6. 6-x, P6 = 1146  
| rowspan=7 | Expected:<br>1. 4-x, P1 = 1145 <br>2. 4-x, P3 = 1046 <br>3. 4-x, P6 = 1139 <br>4. 6-x, P1 = 1146 <br>5. 6-x, P3 = 1046 <br>6. 6-x, P6 = 1146  
| rowspan=7 | [[Image:2016-06-23_6-1_through_6-6.jpg|300px|Hover name]] [[Image:2016-06-23_4-1_through_4-9_no_4-4_or_4-5.jpg|300px|Hover name]]<br>5 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
| rowspan=7 | [[Image:2016-06-23_6-1_through_6-6.jpg|300px|Hover name]]<br>[[Image:2016-06-23_4-1_through_4-9_no_4-4_or_4-5.jpg|300px|Hover name]]<br>5 μL/lane; 1% agarose; [http://openwetware.org/wiki/Image:KAH_Fermentas_GeneRuler_1kbplus.jpg Ladder]
|-
|-
| genomic OR plasmid DNA || 1.0 / 0.5  || 4.0 / 2.0
| genomic OR plasmid DNA || 1.0 / 0.5  || 4.0 / 2.0

Revision as of 07:15, 11 July 2016

Project name <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>

PCR/Agarose Gel Electrophoresis

  • PCR: Candidate cell lines (3 primers + 1 error = 4)
Reagent Vol Mix* (x #) Expected:
1. 4-x, P1 = 1145
2. 4-x, P3 = 1046
3. 4-x, P6 = 1139
4. 6-x, P1 = 1146
5. 6-x, P3 = 1046
6. 6-x, P6 = 1146 		Hover name
Hover name
5 μL/lane; 1% agarose; Ladder
genomic OR plasmid DNA 1.0 / 0.5 4.0 / 2.0
10 μM forward primer 1.0 ---
10 μM reverse primer 1.0 ---
2x GoTaq green 10.0 40.0
dH2O 7.0 / 7.5 28.0 / 30
  20.0 μL 72.0 μL

Note: *Prep one Mix per template

  • For each template, aliquot 18.0 master mix into 3 tubes

Thermal cycler: Labnet - GOTAQ

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


  • Conclusions
  • Top image, top row, 6-1, 6-2, 6-3, P1, 3, and 6 for each.
  • Top image, bottom row, 6-4, 6-5, 6-6, P1, 3, and 6 for each.
  • Bottom image, top row, 4-1, 4-2, 4-3, P1, 3, and 6 for each.
  • Bottom image, bottom row, 4-4, 4-7, 4-9, P1, 3, and 6 for each.



Retrieved from "https://openwetware.org/mediawiki/index.php?title=Haynes_Lab:Notebook/Jan/2016/06/23&oldid=944303"