Difference between revisions of "User:Stuart McKellar/Notebook/Bird Sex Testing/2012/12/01"

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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>}}
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|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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total volume in each is 25μL.
 
total volume in each is 25μL.
  
PCR: 94°C for 2m30s, 35 cycles of: 95°C 30s, 65°C for 30s, 72°C for 30s; final elongation 72°C for 5 mins.
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PCR: 94°C for 2m30s, 35 cycles of: 95°C 30s, 65°C for 30s, 72°C for 30s; final elongation 72°C for 5 mins( actually went 10 mins by accident).
  
 
results to be determined via agar gel electrophoresis on 1% gel stained using gelgreen.
 
results to be determined via agar gel electrophoresis on 1% gel stained using gelgreen.

Latest revision as of 22:18, 26 September 2017

Owwnotebook icon.png Project name Report.pngMain project page
Resultset previous.pngPrevious entry      Next entryResultset next.png

higher annealing temp

Aim: test the p2/p8 and 2550/2178 primers at 65°C.

Method:

Sample 1:

  • A6: p2/p8 primers
  • B6: 2550/2178 primers

Sample 2:

  • C6: p2/p8 primers
  • D6: 2550/2178 primers

Negative control - no DNA (1ul h2O, 2550/2178 primers.)

Master mix (5x):

  • 12.5μL buffer 10x
  • 12.5μL dNTPS
  • 7.5μL MgCl2
  • 62.5 H2O
  • 1μL Taq


  • A6: p2/p8 primers (2.5μL each, 1μL DNA) + 18.5 Master Mix
  • B6: 2550/2178 primers (2.5μL each, 1μL DNA) + 18.5 Master Mix
  • C6: p2/p8 primers (2.5μL each, 1μL DNA) + 18.5 Master Mix
  • D6: 2550/2178 primers (2.5μL each, 1μL DNA) + 18.5 Master Mix

total volume in each is 25μL.

PCR: 94°C for 2m30s, 35 cycles of: 95°C 30s, 65°C for 30s, 72°C for 30s; final elongation 72°C for 5 mins( actually went 10 mins by accident).

results to be determined via agar gel electrophoresis on 1% gel stained using gelgreen.