Difference between revisions of "User:Stuart McKellar/Notebook/Bird Sex Testing/2012/12/02"

From OpenWetWare
(Autocreate 2012/12/02 Entry for User:Stuart_McKellar/Notebook/Bird_Sex_Testing)
 
(Entry title)
Line 6: Line 6:
 
| colspan="2"|
 
| colspan="2"|
 
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
 
<!-- ##### DO NOT edit above this line unless you know what you are doing. ##### -->
==Entry title==
+
==Trying all loris with 65°C PCR - p2/p8 ==
* Insert content here...
+
Aim: To see if at 65°C i can differentiate males from females.
 +
 
 +
Method:
 +
Master mix for all 8 samples plus -ve control(10 units of MM):
 +
*25ul buffer (10x)
 +
*20ul dNTPs
 +
*15ul MgCl2
 +
*1ul Taq
 +
*25ul P2 primer (F)
 +
*25ul P8 primer (R)
 +
*129ul H<sub>2</sub>OH20
 +
 
 +
Tubes labelled as follows:
 +
*A:1SWWBD
 +
*B:2SWWBD
 +
*C:R5#9
 +
*D:R5#51
 +
*E:R5#52
 +
*F:R5#100
 +
*G:Pink Gold
 +
*H:banjo
 +
*-ve: 1ul water
 +
 
 +
Add 1ul of relevant DNA to each tube and 24ul of MM.
 +
 
 +
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.
  
  

Revision as of 20:00, 1 December 2012

Owwnotebook icon.png 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>

Trying all loris with 65°C PCR - p2/p8

Aim: To see if at 65°C i can differentiate males from females.

Method: Master mix for all 8 samples plus -ve control(10 units of MM):

  • 25ul buffer (10x)
  • 20ul dNTPs
  • 15ul MgCl2
  • 1ul Taq
  • 25ul P2 primer (F)
  • 25ul P8 primer (R)
  • 129ul H2OH20

Tubes labelled as follows:

  • A:1SWWBD
  • B:2SWWBD
  • C:R5#9
  • D:R5#51
  • E:R5#52
  • F:R5#100
  • G:Pink Gold
  • H:banjo
  • -ve: 1ul water

Add 1ul of relevant DNA to each tube and 24ul of MM.

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.