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Cloning long fragments


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16 replies to this topic

#16 Heini

Heini

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Posted 14 February 2005 - 03:42 AM

Hi!
Thank you for your reply. I'll try ligation over weekend. I just wonder how much ligase you use for this? And how much of ligation you used for transformation?
(or did you do any inactivation after ligation?) I've also planned to do some temperature cycling -ligation, if this (normal way) doesn't work.

Thank you

#17 claude

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Posted 17 February 2005 - 05:39 AM

Hello Heini!

I use 1ÁL of Ligase as usual in a 20ÁL ligation-mix. After the ligation-reaction I add the whole mix to my bacteria solution. This might not be the proper way, but as it worked several times.....

Keep on trying!

Claude




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