User:Stuart McKellar/Notebook/Bird Sex Testing/2012/12/02

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(Autocreate 2012/12/02 Entry for User:Stuart_McKellar/Notebook/Bird_Sex_Testing)
Current revision (23:00, 1 December 2012) (view source)
(Entry title)
 
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==Entry title==
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==Trying all loris with 65°C PCR - p2/p8 ==
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* Insert content here...
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Aim: To see if at 65°C i can differentiate males from females.
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Method:
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Master mix for all 8 samples plus -ve control(10 units of MM):
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*25ul buffer (10x)
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*20ul dNTPs
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*15ul MgCl2
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*1ul Taq
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*25ul P2 primer (F)
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*25ul P8 primer (R)
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*129ul H<sub>2</sub>OH20
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Tubes labelled as follows:
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*A:1SWWBD
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*B:2SWWBD
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*C:R5#9
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*D:R5#51
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*E:R5#52
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*F:R5#100
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*G:Pink Gold
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*H:banjo
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*-ve: 1ul water
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Add 1ul of relevant DNA to each tube and 24ul of MM.
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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).
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results to be determined via agar gel electrophoresis on 1% gel stained using gelgreen.

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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.



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