Forum: Illumina/Solexa
09-28-2011, 07:53 AM
|
|
Replies: 12
Views: 3,498
I also would recommend against investing on a...
I also would recommend against investing on a GAIIx at this point. The Hiseq has been around for a couple of years already, so the GAIIs sound ancient in terms of the data generated and costs...
|
|
Forum: Sample Prep / Library Generation
09-27-2011, 01:09 PM
|
|
Replies: 26
Views: 31,531
I've never isolated RNA from silkworm, but my...
I've never isolated RNA from silkworm, but my best guess is that it might be similar to the drosophila RNA. I'm sorry that I don't have any RNA profiles in my current lab. But a google search got me...
|
|
Forum: Sample Prep / Library Generation
05-12-2011, 02:10 AM
|
|
Replies: 6
Views: 4,154
|
|
Forum: Sample Prep / Library Generation
05-12-2011, 02:01 AM
|
|
Replies: 5
Views: 3,191
Hi Joe,
I don't work on Roche library preps,...
Hi Joe,
I don't work on Roche library preps, but the problem sounds familiar! In my experience, the Ampure beads are very effective in recovering most of the stuff in the reaction, but when one lets...
|
|
Forum: Sample Prep / Library Generation
05-09-2011, 02:09 PM
|
|
Replies: 5
Views: 4,176
hi Emongs, IMHO, the invitrogen magnet doesn't...
hi Emongs, IMHO, the invitrogen magnet doesn't work any better than other ones. I totally quit using invitrogen magnets and now use magnets which costs nothing compared to the terribly expensive...
|
|
Forum: Sample Prep / Library Generation
05-09-2011, 02:01 PM
|
|
Replies: 6
Views: 4,300
When you say 612 bp, is it the mean insert size...
When you say 612 bp, is it the mean insert size or the size that was gel selected? If it was the selected size, you'd lose around 120 bp for the adapters on both ends which would explain why you get...
|
|
Forum: Sample Prep / Library Generation
05-09-2011, 10:35 AM
|
|
Replies: 5
Views: 4,176
hi bunnymac,
Probably you've figured out the...
hi bunnymac,
Probably you've figured out the issue already, but I just saw the bioanalyzer profile now and realized that it was a RNA Nano. When you say 4/6/8 ug, do you mean total RNA or mRNA? (I...
|
|
Forum: Sample Prep / Library Generation
05-08-2011, 07:05 AM
|
|
Replies: 1
Views: 1,787
Hi Heidi, I would recommend a cleanup of gDNA...
Hi Heidi, I would recommend a cleanup of gDNA (columns, beads or phenol ChCl3) before you do library prep. If I got it right, you want to use the DNA for sequence capture before actual sequencing....
|
|
Forum: Sample Prep / Library Generation
05-08-2011, 06:57 AM
|
|
Replies: 6
Views: 4,300
|
|
Forum: Illumina/Solexa
10-04-2010, 02:20 AM
|
|
Replies: 16
Views: 8,782
|
|
Forum: 454 Pyrosequencing
04-12-2010, 08:30 AM
|
|
Replies: 10
Views: 4,629
Hi Nickloman, I use the NEBnext sample prep kit...
Hi Nickloman, I use the NEBnext sample prep kit for the Illumina library prep. This works quite well for me and Its much cheaper than the Illumina reagents. I don't know about the Roche sample prep...
|
|
Forum: Illumina/Solexa
04-06-2010, 02:05 AM
|
|
Replies: 21
Views: 16,054
I've no clue of why this happens. I tried the...
I've no clue of why this happens. I tried the enzyme again. This time with a longer enzyme inactivation step. I didnt clean up my reaction. But I still have difference between the gel and bioanalyzer...
|
|
Forum: Illumina/Solexa
03-30-2010, 01:24 AM
|
|
Replies: 32
Views: 24,034
I think the cost of the library prep is quite...
I think the cost of the library prep is quite high for the SPRiworks. It would save a lot of hands on time for sure, but I wonder if the throughput that you get for that money is worth it!! The Broad...
|
|
Forum: Illumina/Solexa
03-12-2010, 01:39 AM
|
|
Replies: 21
Views: 16,054
I used the enzyme for just testing the...
I used the enzyme for just testing the fragmentation and was pretty impressed with the fragment size on the gel. It was a focussed smear at 100-600 bp. But when I ran the samples on the bioanalyzer...
|
|
Forum: Sample Prep / Library Generation
03-10-2010, 06:24 AM
|
|
Replies: 23
Views: 18,600
Hi Greigite,
I see a similar band at 130 bp...
Hi Greigite,
I see a similar band at 130 bp in my sequencing library too! Did you figure out why this happens and any way to get rid of this? I tried going down on the adaptor concentration. But...
|
|
Forum: Sample Prep / Library Generation
03-09-2010, 11:48 PM
|
|
Replies: 26
Views: 31,531
Hi Xi wang, I'm sorry if my question was...
Hi Xi wang, I'm sorry if my question was confusing. I meant about the 3 large rRNA subunits. Normally you see 2 of them for mammals and bacteria (In case of drosophila, 2 close peaks at the 18 S and...
|
|
Forum: Sample Prep / Library Generation
03-09-2010, 12:24 AM
|
|
Replies: 26
Views: 31,531
|
|
Forum: Introductions
03-09-2010, 12:14 AM
|
|
Replies: 373
Views: 148,447
|
