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Forum: Sample Prep / Library Generation 07-11-2015, 12:06 PM
Replies: 6
Views: 3,817
Posted By IdahoRAD
When I use these kits, which I really like, I...

When I use these kits, which I really like, I just use the TruSeq adapters. Unless you specifically NEED the NEBNext adapters, why not just use the TruSeq?
Forum: Sample Prep / Library Generation 04-20-2015, 09:54 AM
Replies: 5
Views: 2,750
Posted By IdahoRAD
I think that a clean up is a good idea also. And...

I think that a clean up is a good idea also. And then elute them in a small volume, quantify, then dilute to 50 ng/uL. But I also agree that 50 ng/uL seems pretty high!
Forum: Sample Prep / Library Generation 04-20-2015, 09:49 AM
Replies: 3
Views: 2,280
Posted By IdahoRAD
I have used this same kit, and I am really...

I have used this same kit, and I am really impressed with how it works. I've put in as little as 1 ng and have gotten out over 100 ng. Usually I have a 100-fold increase, with less than 1% being PCR...
Forum: Sample Prep / Library Generation 11-19-2014, 10:20 AM
Replies: 4
Views: 2,270
Posted By IdahoRAD
All I can add is that I have used the Biorupter,...

All I can add is that I have used the Biorupter, by Diagenode. With the biorupter, I have a hard time getting it to shear in the size range that I prefer, it seems to "over shear," so that when I...
Forum: Sample Prep / Library Generation 10-30-2014, 03:47 PM
Replies: 12
Views: 11,455
Posted By IdahoRAD
I have personally left Ampure beads out, ON, on...

I have personally left Ampure beads out, ON, on accident. Paranoid, I thought I couldn't use them, but I did, and everything was just fine!
Forum: Sample Prep / Library Generation 10-14-2014, 01:36 PM
Replies: 10
Views: 2,166
Posted By IdahoRAD
So, you are making sure to only consider the...

So, you are making sure to only consider the amount of DNA, and not the concentration? Changes in elution volumes could make it "look" like the DNA content is increasing.
Forum: Sample Prep / Library Generation 09-18-2014, 08:04 AM
Replies: 4
Views: 1,795
Posted By IdahoRAD
Nucacidhunter-- Thank you for your reply....

Nucacidhunter--

Thank you for your reply. Would you mind explaining your rationale, in the example you gave above? Is it because if we pool equi-molarly, we are doubling the volume and thus the...
Forum: Sample Prep / Library Generation 09-17-2014, 11:21 AM
Replies: 4
Views: 1,795
Posted By IdahoRAD
Annealing adapters

Hello all--

I work on preparing RAD libraries, and have a question regarding annealing adapters.

Suppose my adapters' top and bottom oligos are each at a stock concentration of 100 uM. If I...
Forum: Introductions 01-29-2014, 03:19 PM
Replies: 0
Views: 776
Posted By IdahoRAD
RAD Sequencing in Idaho

Hey all--

I'm a new guy, out in Idaho. I'm working on constructing RAD libraries and I'm totally new to molecular biology. Hope to learn a lot from ya! I'm applying to medical school this year,...
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