Wisconsin/PCR
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< Wisconsin(Difference between revisions)
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| + | ===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 | ||
*5uL 10x ExTaq buffer | *5uL 10x ExTaq buffer | ||
| + | *1uL 10mM dNTPs | ||
| + | *1uL Forward Primer | ||
| + | *1uL Reverse Primer | ||
| + | *1uL Template | ||
| + | *0.5uL ExTaq (add this last) | ||
| + | *2uL MgCl<sub>2</sub> 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] | ||
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]
