Forum: Sample Prep / Library Generation
12-23-2015, 06:05 AM
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Replies: 9
Views: 3,006
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Forum: Sample Prep / Library Generation
12-21-2015, 04:04 AM
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Replies: 0
Views: 1,653
TruSeq nano library changes?
Hello, have anyone noticed their TruSeq nano libraries having a higher yield and/or giving larger size distributions recently?
The new version of the protocol (June 2015) has a change in the...
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Forum: Illumina/Solexa
10-06-2015, 07:12 AM
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Replies: 4
Views: 2,270
HiSeq rapid run overclustering
Hello,
On the Illumina website, it says that the optical raw cluster density for PhiX runs is 850K-1000K clusters/mm2. But, what is the functional range of the instrument in this mode? The GC bias...
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Forum: Sample Prep / Library Generation
07-06-2015, 01:02 AM
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Replies: 3
Views: 1,845
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Forum: Sample Prep / Library Generation
07-03-2015, 03:20 AM
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Replies: 3
Views: 1,845
Blue pippin size selection elution volume
Hello,
Has anyone has tried eluting their TruSeq DNA libraries in a smaller volume than the 40uL stated in the protocol?
We carry out size selection on our TruSeq nano libraries after the PCR...
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Forum: Illumina/Solexa
06-22-2015, 01:11 AM
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Replies: 5
Views: 1,608
No, we don't re-use plates. We clean the...
No, we don't re-use plates. We clean the plate-holder wells with bleach, water and ethanol on recommendation by Kapa once a month. We don't use water to dilute the libraries, we use EB-tween. We put...
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Forum: Illumina/Solexa
06-18-2015, 06:17 AM
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Replies: 5
Views: 1,608
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Forum: Illumina/Solexa
06-18-2015, 01:36 AM
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Replies: 5
Views: 1,608
qPCR issues
Hello, we use Kapa qPCR kits to quantify our libraries before sequencing. Recently, the NTC wells are giving low cq values which means there are contaminants and therefore the data is not reliable....
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Forum: Sample Prep / Library Generation
10-30-2014, 08:42 AM
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Replies: 5
Views: 4,100
Hello,
It's a size selection issue. The...
Hello,
It's a size selection issue. The protocol doesn't need to be altered, just make sure your SP beads are vortexed a lot (I mean a lot!). I usually mix them in between each sample prep as a...
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Forum: Sample Prep / Library Generation
10-29-2014, 02:18 AM
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Replies: 2
Views: 3,905
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Forum: Sample Prep / Library Generation
04-08-2014, 06:19 AM
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Replies: 5
Views: 4,100
Hello, I asked Illumina tech support and they...
Hello, I asked Illumina tech support and they said to try diluting the libraries as the chip looked to be overloaded. I made a 1 in 20 dilution and repeated the chip. An example of a trace is...
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Forum: Sample Prep / Library Generation
04-07-2014, 03:55 AM
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Replies: 5
Views: 4,100
Larger size TruSeq nano prep
Hello,
I followed the protocol for the 550bp size TruSeq nano library preparation but my final libraries were much larger than the expected 670bp average size (see attached pic).
Has anyone else...
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Forum: Sample Prep / Library Generation
10-08-2013, 06:11 AM
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Replies: 0
Views: 1,811
Bimodal distribution in TruSeq libraries
Hi,
I made some control TruSeq genomic gel-free libraries using Ecoli and pooled these together equimolar. This pool then underwent blue pippin size selection at 540bp. The plot after duplicate...
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Forum: Sample Prep / Library Generation
09-10-2013, 06:54 AM
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Replies: 2
Views: 2,771
Extra peak after TruSeq DNA library prep
Hello,
We made library preps following the standard TruSeq DNA library prep method, beginning with 3ug gDNA and took this through the steps shearing-end repair-A tailing-ligation-size selection-PCR....
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Forum: Sample Prep / Library Generation
06-17-2013, 06:56 AM
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Replies: 7
Views: 4,718
Hi,
Thanks for your reply. 2 minutes just...
Hi,
Thanks for your reply. 2 minutes just doesn't seem enough. Is there not any ethanol carryover this way? I keep an eye on the beads and usually 10-15 minutes is fine so that the beads appear matt...
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Forum: Sample Prep / Library Generation
06-17-2013, 06:54 AM
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Replies: 0
Views: 2,450
Size selection for TruSeq DNA libraries
Hello,
When is the best time for carrying out size selection of TruSeq DNA libraries - pre or post PCR? What kind of adapters are labs generally using?
We've been doing a method of using a paired...
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Forum: Sample Prep / Library Generation
06-17-2013, 06:47 AM
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Replies: 7
Views: 4,718
Ampure bead clean up
Hello,
How much drying time do you normally allow when using ampure beads XP to clean up your library preps?
I've been using the standard 15 minutes as mentioned in the TruSeq DNA library prep...
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