User:Stuart McKellar/Notebook/Bird Sex Testing/2012/12/02: Difference between revisions
From OpenWetWare
(Autocreate 2012/12/02 Entry for User:Stuart_McKellar/Notebook/Bird_Sex_Testing) |
|||
| 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. ##### --> | ||
== | ==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 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
 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> </html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html> |
Trying all loris with 65°C PCR - p2/p8Aim: 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):
Tubes labelled as follows:
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.
| |
