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Forum: Sample Prep / Library Generation 03-05-2014, 01:58 AM
Replies: 3
Views: 1,277
Posted By amazonic9
Hi Chipper, thanks for you answer. ...

Hi Chipper,

thanks for you answer.

Yesteday we did a test with the NebNext Ultra kit, starting with 1 ng of fragmented DNA, and at the point of the PCR, we tested three polymerases: NebNext...
Forum: Sample Prep / Library Generation 03-03-2014, 02:07 AM
Replies: 1
Views: 1,038
Posted By amazonic9
Sorry the thread was duplicated somehow. If...

Sorry the thread was duplicated somehow. If someone can delete one of them...?

Thanks
Forum: Sample Prep / Library Generation 03-03-2014, 01:13 AM
Replies: 1
Views: 1,038
Posted By amazonic9
Polymerase for ChIP-seq. Which do you use?

Dear all,

I would like to know which polymerase do you use for your ChIP-seq libraries?

We use the phusion polymerase, but I would like to know your opinion about the other polymerases, and if...
Forum: Sample Prep / Library Generation 03-03-2014, 01:12 AM
Replies: 3
Views: 1,277
Posted By amazonic9
Polymerase for ChIP-seq. Which do you use?

Dear all,

I would like to know which polymerase do you use for your ChIP-seq libraries?

We use the phusion polymerase, but I would like to know your opinion about the other polimerases, and if...
Forum: Sample Prep / Library Generation 12-05-2012, 02:56 AM
Replies: 2
Views: 2,429
Posted By amazonic9
Yes, that is what I thought, that it was a matter...

Yes, that is what I thought, that it was a matter of size (small sizes). But as I have seen people doing selection on acrylamida gels at 300bp for example (with this directional mRNA protocol using...
Forum: Sample Prep / Library Generation 12-04-2012, 08:17 AM
Replies: 2
Views: 2,429
Posted By amazonic9
Size selection: agarose gel / acrylamide gel

Hello everybody,

Maybe I am going to ask something very basic, but I haven't found the answer on the internet (probably I did the wrong queries)...

Could anyone tell me why we usually make size...
Forum: Illumina/Solexa 11-27-2012, 01:42 AM
Replies: 3
Views: 1,992
Posted By amazonic9
Hi! I guess it should work, as we have...

Hi!

I guess it should work, as we have received Truseq RNA kits with the Truseq DNA PCR box... so, either Illumina had a problem with the shipment and we received something we were not supposed to...
Forum: Sample Prep / Library Generation 01-30-2012, 06:29 AM
Replies: 16
Views: 17,355
Posted By amazonic9
Again, thank you for your answers. I think this...

Again, thank you for your answers. I think this sentence could be the key "Sometimes the issue is that one activity works under one set of conditions while another is optimum under a different set."
Forum: Sample Prep / Library Generation 01-25-2012, 07:55 AM
Replies: 16
Views: 17,355
Posted By amazonic9
Thank you phillip for the answer. That makes...

Thank you phillip for the answer. That makes sense to me. But in the particular case that I am using this phosphatase + PNK treatment, do not ligate a 3' adapter before the treatment with PNK. The...
Forum: Sample Prep / Library Generation 01-23-2012, 08:15 AM
Replies: 16
Views: 17,355
Posted By amazonic9
Hello all, Can anyone tell me why in some...

Hello all,

Can anyone tell me why in some protocols (Directional mRNA-seq) one has to treat with a phosphatase (that removes de 5' phosphate) and then with T4 PNK after RNA fragmentation? I mean,...
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