Talk:DNA ligation: Difference between revisions
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==Insert to Vector ratios== | ==Insert to Vector ratios== | ||
Hi, new to this wiki but not to wiki's. I notice that a formula was used for a 6:1 insert:vector ratio at the top of the page but later it talks about the appropraite range is 1:1 to 5:1, this seems like a contradiction. Under what conditions does one use the 6:1 ratio. | Hi, new to this wiki but not to wiki's. I notice that a formula was used for a 6:1 insert:vector ratio at the top of the page but later it talks about the appropraite range is 1:1 to 5:1, this seems like a contradiction. Under what conditions does one use the 6:1 ratio. This page needs at least some discussion of how to ligate vectors to inserts where both are reasonably long. Phil 18:22, 6 September 2007 (EDT). | ||
Revision as of 22:24, 6 September 2007
General ligation protocols are fine for protruding sticky end ligations but often fail with blunt ends or recessed sticky ends. For blunt ends, higher enzyme concentrations are recommended along with considerably lower total DNA concentrations (cf. Bercovich et al., BioTechniques 12(2), 190-193 (1992)). If there is someone around with a good working protocol for ligation of blunt and of protruding ends, it would be much appreciated. {MDolinar, July 25, 2007}
Insert to Vector ratios
Hi, new to this wiki but not to wiki's. I notice that a formula was used for a 6:1 insert:vector ratio at the top of the page but later it talks about the appropraite range is 1:1 to 5:1, this seems like a contradiction. Under what conditions does one use the 6:1 ratio. This page needs at least some discussion of how to ligate vectors to inserts where both are reasonably long. Phil 18:22, 6 September 2007 (EDT).
