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help ,blunt end ligation

blunt end ligation

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4 replies to this topic

#1 pppyeah

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Posted 03 September 2012 - 02:20 AM

hi,

I do blunt end ligation over night by seting the temp 12°C as it should be 16°C。

and now i increase the temp to 16°C.

did this makes any different to the result?

thanks thanks

#2 Inbox

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Posted 03 September 2012 - 02:26 AM

You may get colonies but may beless ligation efficiency.

#3 pppyeah

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Posted 03 September 2012 - 03:13 AM

You may get colonies but may beless ligation efficiency.


thanks, can i do something now to improve?
MY plan is keep it 16 for another 5 hour

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Posted 03 September 2012 - 03:44 AM

No I don't think there is much you can do when temperature is not set at desired. But Don't worry you should get colonies at 12 0C incubation too.
People usually do 4 0C overnight incubation. Its may be better you directly try transformation now.

#5 pppyeah

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Posted 03 September 2012 - 05:16 AM

No I don't think there is much you can do when temperature is not set at desired. But Don't worry you should get colonies at 12 0C incubation too.
People usually do 4 0C overnight incubation. Its may be better you directly try transformation now.

actually i am not doing transformation,this is just a step in my microarray preparation.

thanks





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