Addition of 3' A overhangs to PCR products: Difference between revisions
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===Reagents=== | ===Reagents=== | ||
*Purified PCR product | |||
*dATP | |||
*PCR buffer with Mg | |||
*''Taq'' DNA Polymerase | |||
*ddH<sub>2</sub>O | |||
===Equipment=== | ===Equipment=== | ||
*Thermocycler or Heat block equilibrated at 72°C | |||
*Spectrophotometer | |||
==Procedure== | ==Procedure== | ||
Revision as of 20:41, 18 October 2008
Curators
Mohsen Hosseini
Abstract
If PCR is performed using a proofreading DNA polymerase, such as Pfu DNA polymerase, the product will have blunt ends. Taq DNA polymerase catalyzes the non-template directed addition of an adenine residue to the 3´-end of both strands of DNA molecules to make it suitable for TA cloning.
Materials
Reagents
- Purified PCR product
- dATP
- PCR buffer with Mg
- Taq DNA Polymerase
- ddH2O
Equipment
- Thermocycler or Heat block equilibrated at 72°C
- Spectrophotometer
Procedure
Critical steps
Troubleshooting
Notes
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