Wisconsin/PCR
From 2007.igem.org
< Wisconsin(Difference between revisions)
| Line 1: | Line 1: | ||
| + | ===Primer Stock=== | ||
| + | First check primer concentration using spectrophotometer... then prepare stock of 500uL 10uM. | ||
| + | |||
===Basic Non-Sequencing PCR=== | ===Basic Non-Sequencing PCR=== | ||
*38.5uL H<sub>2</sub>O | *38.5uL H<sub>2</sub>O | ||
Latest revision as of 19:41, 4 July 2007
Primer Stock
First check primer concentration using spectrophotometer... then prepare stock of 500uL 10uM.
Basic Non-Sequencing PCR
- 38.5uL H2O
- 5uL 10x ExTaq buffer
- 1uL 10mM dNTPs
- 1uL Forward Primer
- 1uL Reverse Primer
- 1uL Template
- 0.5uL ExTaq (add this last)
- 2uL MgCl2 25mM
Run on Ansari PCR machine CDC-02
PCR Purification
Follow instruction in the [http://www1.qiagen.com/literature/protocols/QIAquickSpin.aspx QIAquick Spin Handbook]
