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

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|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
|style="background-color: #EEE"|[[Image:owwnotebook_icon.png|128px]]<span style="font-size:22px;"> Project name</span>
|style="background-color: #F2F2F2" align="center"|<html><img src="/images/9/94/Report.png" border="0" /></html> [[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>[[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]]<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>}}
|style="background-color: #F2F2F2" align="center"|[[File:Report.png|frameless|link={{#sub:{{FULLPAGENAME}}|0|-11}}]][[{{#sub:{{FULLPAGENAME}}|0|-11}}|Main project page]]<br />{{#if:{{#lnpreventry:{{FULLPAGENAME}}}}|[[File:Resultset_previous.png|frameless|link={{#lnpreventry:{{FULLPAGENAME}}}}]][[{{#lnpreventry:{{FULLPAGENAME}}}}{{!}}Previous entry]]&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;}}{{#if:{{#lnnextentry:{{FULLPAGENAME}}}}|[[{{#lnnextentry:{{FULLPAGENAME}}}}{{!}}Next entry]][[File:Resultset_next.png|frameless|link={{#lnnextentry:{{FULLPAGENAME}}}}]]}}
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<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
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==Entry title==
==PCR/Agarose Gel Electrophoresis==
* Insert content here...
*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. 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]]<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
|-
| 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 ||  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.




Latest revision as of 01:49, 27 September 2017

Project name Main project page
Previous entry      

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.



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