Melbourne/Lab BL Notebook/PCR1

From 2007.igem.org

(Difference between revisions)
Line 5: Line 5:
5ul Enhancer
5ul Enhancer
 +
0.6ul dNTPs (25mM stock)
0.6ul dNTPs (25mM stock)
 +
2.5ul MgSO4 (Supplied in PCR kit)
2.5ul MgSO4 (Supplied in PCR kit)
 +
1.5ul Primer I (10uM)
1.5ul Primer I (10uM)
-
*1.5ul Primer II (10uM)
 
-
*1ul Template ([[Melbourne/pJS010|pJS010]])
 
-
*0.4ul Pfx Platinum (Invitrogen)
 
-
*32.5ul ddH<sub>2</sub>O
 
 +
1.5ul Primer II (10uM)
 +
 +
1ul Template ([[Melbourne/pJS010|pJS010]])
 +
 +
0.4ul Pfx Platinum (Invitrogen)
 +
 +
32.5ul ddH<sub>2</sub>O
--
--
-
*50ul Total
+
50ul Total

Revision as of 14:47, 10 October 2007

=Protocol for PCR reactions A~G= (and -ve control) amplifying the photoreceptor and transmembrane domains of NpSopII-NpHtrII.

PCR mix

5ul 10x buffer\\

5ul Enhancer

0.6ul dNTPs (25mM stock)

2.5ul MgSO4 (Supplied in PCR kit)

1.5ul Primer I (10uM)

1.5ul Primer II (10uM)

1ul Template (pJS010)

0.4ul Pfx Platinum (Invitrogen)

32.5ul ddH2O -- 50ul Total