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Old 06-09-2015, 03:31 AM   #16
kobeho24
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Location: HKUST, Hong Kong

Join Date: Apr 2015
Posts: 32
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Quote:
Originally Posted by Simone78 View Post
Have you tried to do the assembly in solution as we describe in the paper? Just elute the tn5 from the column and add the oligos in equimolar conc with the Tn5 --> incubate 37 degrees --> ready to use. If that doesn´t work then you might have an issue with the Tn5.
Best,
Simone
Hi Simone,
I tried to produce the Tnp once again with the C3013 lately. But the strain seemed to reach plateau phase after the A600=1.3. No matter there is IPTG induction or not, same thing happened. However, according to your protocol, the A600 should reach 2.1 after IPTG induction for around 4h. The protein solution is still being dialyzed, I will test if there is any activity after that in any case. But the strain seems weird on our hands. Maybe you can give me some advices. Thanks!

Cheers,
Gary
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