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Forum: Illumina/Solexa 05-31-2017, 09:23 AM
Replies: 23
Views: 8,391
Posted By jlei_face
As you have probably already seen, Illumina will...

As you have probably already seen, Illumina will be working with IDT to create these UDI's. I know that IDT already has 96 unique indices, 8 nt long, as I have used them in the past. Instead of...
Forum: Sample Prep / Library Generation 10-05-2015, 03:53 PM
Replies: 3
Views: 1,275
Posted By jlei_face
Hey EGarf, My facility is very high throughput...

Hey EGarf,
My facility is very high throughput so we have moved away from qPCR given both the cost and the time. We do picogreen (Quant-iT) paired with LabChip (similar to BioAnalyzer). Our...
Forum: Illumina/Solexa 08-25-2015, 08:04 PM
Replies: 2
Views: 2,228
Posted By jlei_face
Thanks for sharing this data. I'd just like to...

Thanks for sharing this data.
I'd just like to add that concentrations of PhiX vary from lot to lot so 8nM wouldn't necessarily be applicable to every tube.
Forum: Sample Prep / Library Generation 08-22-2015, 06:37 AM
Replies: 6
Views: 2,839
Posted By jlei_face
I know this reply is super late but since it...

I know this reply is super late but since it seems there are other posters with similar issues I'll put in my two cents:
When doing a library prep for first few times, it is recommended that you...
Forum: Illumina/Solexa 08-22-2015, 06:22 AM
Replies: 3
Views: 2,742
Posted By jlei_face
What is the end goal of this? Taxonomic...

What is the end goal of this? Taxonomic assignment or de novo reconstruction of the samples? How many samples do you need to sequence?
Additionally, I would recommend skipping the bead normalization...
Forum: Sample Prep / Library Generation 08-22-2015, 05:37 AM
Replies: 3
Views: 2,489
Posted By jlei_face
In general, I would say the kit is still good....

In general, I would say the kit is still good. However, the fact that the reagents have gone through a freeze-thaw is something that may contribute to degradation in addition to time. Usually...
Forum: Illumina/Solexa 12-10-2014, 06:44 AM
Replies: 3
Views: 1,826
Posted By jlei_face
The ligation of adapters (which contain your...

The ligation of adapters (which contain your index) occurs right before amplification so as long as you have enough material/high enough library yield for clustering you don't necessarily need to...
Forum: Sample Prep / Library Generation 12-09-2014, 04:35 PM
Replies: 3
Views: 1,517
Posted By jlei_face
Here's a good link: ...

Here's a good link:
http://www.cureffi.org/2012/12/11/how-pcr-duplicates-arise-in-next-generation-sequencing/
Forum: Illumina/Solexa 12-09-2014, 04:27 AM
Replies: 6
Views: 3,311
Posted By jlei_face
AStretton, I've been out of that lab for some...

AStretton,
I've been out of that lab for some time but I believe our optimum temperature was right at RT (25degrees C) or slightly above. The lab that contained the BA also contained other...
Forum: Illumina/Solexa 12-06-2014, 06:26 AM
Replies: 6
Views: 3,311
Posted By jlei_face
Sorry for the late post but I experienced this...

Sorry for the late post but I experienced this problem before also. For my lab, it turns out the ambient temperature was too cold. We ended up creating an enclosure for the BA which kept the air...
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