Knight:TOPO TA cloning: Difference between revisions
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==Notes== | ==Notes== | ||
*[[Tom Knight]] claims you can cut the reaction volume in half and it will still work fine. | *[[Tom Knight]] claims you can cut the reaction volume in half and it will still work fine. | ||
*Use fresh PCR product for best results. (The A overhang tends to get chewed back over time) | *Use fresh PCR product for best results. (The A overhang tends to get chewed back over time ... even using day old PCR product can reduce efficiency.) | ||
*Your primers should NOT have 5' phosphates. They interfere with topoisomerase I. | *Your primers should NOT have 5' phosphates. They interfere with topoisomerase I. | ||
Revision as of 14:30, 29 September 2005
Notes
- Tom Knight claims you can cut the reaction volume in half and it will still work fine.
- Use fresh PCR product for best results. (The A overhang tends to get chewed back over time ... even using day old PCR product can reduce efficiency.)
- Your primers should NOT have 5' phosphates. They interfere with topoisomerase I.
