Haynes Lab:Notebook/Jan/2016/06/15: Difference between revisions

From OpenWetWare
Jump to navigationJump to search
Line 8: Line 8:
==Genomic DNA Extraction, PCR, Agarose Gel Verification==
==Genomic DNA Extraction, PCR, Agarose Gel Verification==
* Finished extracting genomic DNA from 4-1, 4-2, 4-3, 6-1, 6-2, 6-5, 6-6.
* Finished extracting genomic DNA from 4-1, 4-2, 4-3, 6-1, 6-2, 6-5, 6-6.
PCR: Candidate cell lines (3 primers + 1 error = 4)
{| {{table}} cellspacing="3" <!-- PCR rxn table -->
| bgcolor=#cfcfcf | Reagent
| bgcolor=#cfcfcf | Vol
| bgcolor=#cfcfcf | Mix* (x #)
| rowspan=7 | Expected:<br>1. CMV = 588
| rowspan=7 | [[Image:somegel.jpg|150px|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
|-
| 10 μM forward primer    || 1.0  || ---
|-
| 10 μM reverse primer  || 1.0  || ---
|-
| 2x GoTaq green    || 10.0 || 40.0
|-
| dH<sub>2</sub>O    || 7.0 / 7.5  || 28.0 / 30
|-
| &nbsp;            || 20.0 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




Revision as of 12:22, 15 June 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>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

Genomic DNA Extraction, PCR, Agarose Gel Verification

  • Finished extracting genomic DNA from 4-1, 4-2, 4-3, 6-1, 6-2, 6-5, 6-6.

PCR: Candidate cell lines (3 primers + 1 error = 4)

Reagent Vol Mix* (x #) Expected:
1. CMV = 588 		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 ---

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



Retrieved from "https://openwetware.org/mediawiki/index.php?title=Haynes_Lab:Notebook/Jan/2016/06/15&oldid=943194"