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Forum: Sample Prep / Library Generation 02-09-2016, 06:17 AM
Replies: 289
Views: 154,271
Posted By daniel007
Hi Simone, With reference to your trials of...

Hi Simone,

With reference to your trials of 2ul tagmentation, did you have any luck reducing the size of your libraries? Even with 250pg input, I find that my final libraries are average 600bp on...
Forum: Sample Prep / Library Generation 08-19-2015, 01:32 PM
Replies: 289
Views: 154,271
Posted By daniel007
Hi Simone, You mentioned scaling down the...

Hi Simone,

You mentioned scaling down the tagmentation reaction to 2ul, then having a 5ul final volume after PCR: do you use your own Tn5 for this, and is 5ul the PCR volume (I've never done a PCR...
Forum: Sample Prep / Library Generation 06-15-2015, 05:18 AM
Replies: 33
Views: 28,431
Posted By daniel007
Did you find it difficult to achieve the...

Did you find it difficult to achieve the onto-column flow rates reported in the Genome Research paper? Is there much difference between the paper and the protocol you received from Sandberg lab? I...
Forum: Sample Prep / Library Generation 06-14-2015, 11:40 AM
Replies: 33
Views: 28,431
Posted By daniel007
I received the plasmid from Addgene, indeed. May...

I received the plasmid from Addgene, indeed. May I ask what equipment you use for the purification - i.e. AKTA etc? Have you tried both on-column and in solution assembly?
Forum: Sample Prep / Library Generation 06-13-2015, 08:54 AM
Replies: 33
Views: 28,431
Posted By daniel007
Hi all, I'm also trying to purify Tn5 using the...

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...
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