Quote:
Originally Posted by daniel007
Hi all,
I'm also trying to purify Tn5 using the pTXB1 plasmid, and I also don't have transpososome activity when assembled in solution with oligos. Does anyone have a slightly more detailed protocol (I'm a molecular biologist playing at protein biochemistry...) - especially regarding how to carry out the column steps, recommended equipment etc? I think my attempt was too drawn out in time (even though I did most of it in a cold room at 4C) and this may have led to incorrect folding/degradation. Any advice much appreciated!
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Up till now, I've already had seveal attempts. Nothing's wrong except for the activity. And now I am kinda puzzled. May I ask where did you get the pTXB1-Tn5 plasmid? Is it from Addgene?