Melbourne/Lab BL Notebook/PCR1

From 2007.igem.org

(Difference between revisions)
Line 24: Line 24:
32.5ul ddH<sub>2</sub>O
32.5ul ddH<sub>2</sub>O
-
|94°C  5'
+
|94°C - 5'
-
94°C   30''
+
94°C - 30''
-
59°C   30''
+
59°C - 30''
-
68°C   35''
+
68°C - 35''
-
68°C   1.5'
+
68°C - 1.5'
4°C forever
4°C forever

Revision as of 15:01, 10 October 2007

Protocol for PCR reactions A~G

For amplifying the photoreceptor and transmembrane domains of NpSopII-NpHtrII.

PCR mix PCR program
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

94°C - 5'

94°C - 30

59°C - 30

68°C - 35

68°C - 1.5'

4°C forever

50ul Total

|-

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 | |-