SEQanswers

Go Back   SEQanswers > Search Forums


Showing results 1 to 17 of 17
Search took 0.00 seconds.
Search: Posts Made By: SarahNGS
Forum: Sample Prep / Library Generation 03-25-2013, 08:59 AM
Replies: 6
Views: 6,363
Posted By SarahNGS
Invitrogen makes a PicoGreen kit containing a...

Invitrogen makes a PicoGreen kit containing a standard:...
Forum: Sample Prep / Library Generation 01-08-2013, 07:16 AM
Replies: 49
Views: 70,862
Posted By SarahNGS
What kind of magnet are you using?

What kind of magnet are you using?
Forum: Sample Prep / Library Generation 01-07-2013, 07:10 AM
Replies: 9
Views: 7,357
Posted By SarahNGS
My money's on too much DNA. Try quantifying...

My money's on too much DNA. Try quantifying your ladder via fluorescence. Or just dilute the DNA 10x and repeat the experiment.
Forum: Sample Prep / Library Generation 11-05-2012, 07:29 AM
Replies: 1
Views: 3,218
Posted By SarahNGS
PCR clean-up for sequencing

Alpineda,
Given how much money you are about to spend sequencing these samples, and the potential precious nature of them, I would not risk it. Do the magnetic bead clean-up! If you do it in a...
Forum: Sample Prep / Library Generation 10-15-2012, 07:22 AM
Replies: 6
Views: 6,363
Posted By SarahNGS
yes you want to look for a "fluorescence plate...

yes you want to look for a "fluorescence plate reader". PerkinElmer makes several including the EnSpire and Victor. They both work great for Pico Green.
Forum: Sample Prep / Library Generation 08-27-2012, 06:08 AM
Replies: 5
Views: 7,549
Posted By SarahNGS
underdrying beads

Not getting all of the ethanol off the beads can also significantly reduce yields. make sure that you see small cracks and that the color lightens up. I have not had a problem with overdrying.
Forum: Sample Prep / Library Generation 07-23-2012, 10:03 AM
Replies: 6
Views: 2,561
Posted By SarahNGS
shearing differences

I'm told (but have no direct experience) that the "tagmentation" step, which uses enzymatic fragmentation via an engineered transposase, has some non-randomness that could potentially result in...
Forum: Ion Torrent 04-16-2012, 06:27 AM
Replies: 18
Views: 9,710
Posted By SarahNGS
not an either/or

Actually pippen prep is for what I call "preparative scale" analysis. this machine and others like it (LabChip XT for example) are designed to recover the sample. It is the automatic equivalent of...
Forum: Sample Prep / Library Generation 04-02-2012, 06:11 AM
Replies: 13
Views: 3,203
Posted By SarahNGS
Thumbs up ditto, its about clean DNA, which most DNA...

ditto, its about clean DNA, which most DNA clean-up kits wil give you.
Forum: Sample Prep / Library Generation 04-02-2012, 06:07 AM
Replies: 7
Views: 8,281
Posted By SarahNGS
DNA quantitation and pooling

Let's hope you're not really submitting DNA of this length...
Fluorimetry methods like Quant It/ Pico Green measure DNA concentration by mass. so if you have a 200 bp PCR product and a 400 bp...
Forum: Ion Torrent 03-26-2012, 07:43 AM
Replies: 10
Views: 11,551
Posted By SarahNGS
Question AmpliSeq Cancer Panel

Does anyone here have experience with the Ion Torrent Ampliseq Cancer Panel? I have a couple of questions-
How satisfied are you with the results?
How many samples are you running per batch?
Forum: Sample Prep / Library Generation 12-27-2011, 06:34 AM
Replies: 49
Views: 70,862
Posted By SarahNGS
re-using AMPure beads

There was a paper published this year by the Broad institute, and they re-use the SPRI beads. :)

http://www.ncbi.nlm.nih.gov/pubmed/21205303

BTW I advise anyone doing NGS to find and read all...
Forum: Sample Prep / Library Generation 11-29-2011, 06:06 AM
Replies: 12
Views: 12,144
Posted By SarahNGS
You may want to consider an alternate means of...

You may want to consider an alternate means of quantitation. NanoDrop quantitation, as stated above, suffers from contaminants. A dsDNA specific method like picogreen will tell you how much DNA you...
Forum: Sample Prep / Library Generation 11-28-2011, 06:43 AM
Replies: 10
Views: 6,512
Posted By SarahNGS
I don't agree that there is "no good solution"...

I don't agree that there is "no good solution" for this. Again it depends on the degree of automation that you want and are willing to pay for. For complete automation yes you would need to...
Forum: 454 Pyrosequencing 11-14-2011, 04:11 AM
Replies: 2
Views: 1,916
Posted By SarahNGS
454 automation options

It is definitely worth looking into automation, and based on the workflow I'd try to automate the library preparation first. The REM e only automates the post-PCR enrichment step and has to be...
Forum: Sample Prep / Library Generation 11-14-2011, 04:04 AM
Replies: 10
Views: 6,512
Posted By SarahNGS
'fully automated' NGS library prep

It is possible with more flexible robots, such as the JANUS form PerkinElmer, to have a more fully automated yet flexible platform for NGS library prep. For example you can integrate a plate sealer...
Forum: 454 Pyrosequencing 11-07-2011, 01:47 PM
Replies: 2
Views: 1,662
Posted By SarahNGS
Question SPRI for 8kb and/or 20kb PE libraries?

Hello,
I have seen some publications and threads about using SPRI beads instead of a gel to purify fragments for the Roche/454 3 kb paired-end prep. Is anyone using SPRI also for the 8 or 20kb...
Showing results 1 to 17 of 17

 


All times are GMT -8. The time now is 02:19 PM.


Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2020, vBulletin Solutions, Inc.
Single Sign On provided by vBSSO