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Forum: Sample Prep / Library Generation 10-11-2016, 07:25 AM
Replies: 2
Views: 2,024
Posted By Manna
I've had great luck using these: Sera-mag...

I've had great luck using these:
Sera-mag SpeedBeads (Fisher#09-981-123)
You basically just wash them, add your own PEG solution, and bingo! you've made AMPure beads for almost zero dollars.
Forum: Sample Prep / Library Generation 10-11-2016, 07:15 AM
Replies: 2
Views: 907
Posted By Manna
Probably not, but you can call illumina, tell...

Probably not, but you can call illumina, tell them what happened, and see if they'll send a replacement. Or possibly, just buy that component.
Forum: Sample Prep / Library Generation 10-06-2016, 07:47 AM
Replies: 20
Views: 9,119
Posted By Manna
I'm using it, though for a DNA-modification, not...

I'm using it, though for a DNA-modification, not a regular ChIP against some DNA-bound protein. Does anyone know the sequences of Active Motif's ridiculously priced indexing primers? Their tech...
Forum: Sample Prep / Library Generation 02-09-2015, 07:03 AM
Replies: 13
Views: 10,667
Posted By Manna
OK, thanks. Looks like I'm in for one more...

OK, thanks. Looks like I'm in for one more bottle of AMPure.
Now... to tell the boss or not to tell him?
Forum: Sample Prep / Library Generation 02-09-2015, 06:39 AM
Replies: 13
Views: 10,667
Posted By Manna
I ordered these SpeedBeads Jan 29, and they...

I ordered these SpeedBeads Jan 29, and they haven't shown up yet (Feb 9), so I called Fisher. Their rep said they were a custom item, and it may take 1-3 months before I actually get them. Is this...
Forum: Sample Prep / Library Generation 01-08-2015, 07:26 AM
Replies: 4
Views: 1,755
Posted By Manna
Also, keep in mind that Covaris has different...

Also, keep in mind that Covaris has different tubes for different applications. I haven't looked into it specifically for your wants, but you might find a different tube type more accommodating for...
Forum: Sample Prep / Library Generation 01-08-2015, 07:23 AM
Replies: 5
Views: 3,103
Posted By Manna
With the TruSeq kit, I typically finish with 30...

With the TruSeq kit, I typically finish with 30 ÁL of library at a concentration of around 10-20 ng/Ál, sometimes as much as 40. Anything less than 5 ng/Ál is suspect, meaning, it isn't likely to...
Forum: Sample Prep / Library Generation 01-08-2015, 07:15 AM
Replies: 16
Views: 5,932
Posted By Manna
If you don't have a bioanalyzer, and you don't...

If you don't have a bioanalyzer, and you don't see anything on a gel, I would suggest a fluorescent nucleic acid detection set up such as Qubit.
Forum: Sample Prep / Library Generation 08-11-2014, 02:18 PM
Replies: 5
Views: 2,296
Posted By Manna
I have encountered this problem. Once the beads...

I have encountered this problem. Once the beads are pulled to the side of the tube, remove the original supernatant / binding buffer, but leave some behind. Leave behind whatever you need to so as...
Forum: Sample Prep / Library Generation 08-11-2014, 01:47 PM
Replies: 2
Views: 4,575
Posted By Manna
The internets don't seem to have any pictures of...

The internets don't seem to have any pictures of what this ladder looks like (Illumina SRA ladder). I have this ladder in my freezer, and I just ran a gel with it, not terribly concerned about...
Forum: Sample Prep / Library Generation 05-22-2013, 12:16 PM
Replies: 0
Views: 1,013
Posted By Manna
real time PCR for library quant?

I am working to make some small RNA sequencing libraries, and have just started using real time PCR to quantify them. The final steps of my library construction are to do 12 PCR cycles, then gel...
Forum: Sample Prep / Library Generation 05-22-2013, 11:58 AM
Replies: 3
Views: 2,283
Posted By Manna
It works totally fine. I've done it. The two...

It works totally fine. I've done it. The two kits a indistinguishable once you have clean dsDNA. The primers and reaction mixes are at the same concentrations.
Forum: Sample Prep / Library Generation 05-22-2013, 11:57 AM
Replies: 1
Views: 1,391
Posted By Manna
I would spin them down and send them dry. RNA is...

I would spin them down and send them dry. RNA is pretty stable this way at a broad range of temperatures. Make sure your recipient knows they're coming that way.
Forum: Sample Prep / Library Generation 05-22-2013, 11:50 AM
Replies: 2
Views: 1,322
Posted By Manna
If you need both RNA & DNA from the same prep,...

If you need both RNA & DNA from the same prep, then split your prep in half. Treat one half with DNase, then run it over a column (qiagen/zymo whatever) to clean up. Base hydrolyze the other half,...
Forum: Sample Prep / Library Generation 02-21-2013, 12:05 PM
Replies: 10
Views: 4,100
Posted By Manna
Best Way to make Mate-Pair library?

What do people think is the best way to make a mate-pair library? I need to do some with 3 and/or 6 kb inserts and probably at least one with a larger 10kb insert. Are there any consistent problems...
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