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Forum: Illumina/Solexa 10-16-2019, 06:20 AM
Replies: 0
Views: 945
Posted By acockburn
Question NextSeq underclustering

Has anyone had issues with underclustering on the Illumina NextSeq 500/550? We have been having issues all summer. Even increasing input 3 times our normal has not caused overclustering. We...
Forum: Illumina/Solexa 05-05-2017, 04:39 AM
Replies: 6
Views: 2,940
Posted By acockburn
Sequal prep ...

Sequal prep
https://www.thermofisher.com/order/catalog/product/A1051001

This is how we normalize.
Forum: Sample Prep / Library Generation 11-17-2015, 05:56 AM
Replies: 0
Views: 1,810
Posted By acockburn
Smile Alternatives to SequalPrep

Hi,

I am looking for alternatives to SequalPrep as they are on backorder for a few months and we are in busy mode.

Thanks
Forum: Illumina/Solexa 05-27-2015, 08:41 AM
Replies: 2
Views: 1,472
Posted By acockburn
I use the Kozich et al. primer set. I have...

I use the Kozich et al. primer set. I have never seen anything like this.

Could you give more info on the library generation?

Lately the reagents for the MiSeq has been crap and my Q30 have...
Forum: Sample Prep / Library Generation 02-24-2015, 07:24 AM
Replies: 19
Views: 6,885
Posted By acockburn
Assuming you want to load 10 pM, 1% spike in of...

Assuming you want to load 10 pM, 1% spike in of PhiX and using the above subnanomolar protocol.

Add the following --- Volume (uL)
0.15 nM sample DNA --- 52
0.2...
Forum: Sample Prep / Library Generation 02-22-2015, 06:59 AM
Replies: 19
Views: 6,885
Posted By acockburn
Are you trying to go lower then 100 pM?

Are you trying to go lower then 100 pM?
Forum: Sample Prep / Library Generation 02-22-2015, 06:58 AM
Replies: 19
Views: 6,885
Posted By acockburn
I usually make 700 uL and load 600 uL. I have...

I usually make 700 uL and load 600 uL. I have never tried less.
Forum: Sample Prep / Library Generation 12-18-2014, 12:39 PM
Replies: 3
Views: 2,546
Posted By acockburn
EpMotion - NEBnext ultra program

Was wondering if there is a program out there for genomic library prep on the EpMotion. I know they have the TruSeq RNA one's, but I am more looking for DNA and hoping specfically for NEBNext Ultra....
Forum: Sample Prep / Library Generation 12-10-2014, 08:27 AM
Replies: 19
Views: 6,885
Posted By acockburn
For subnanomolar libraries we use the protocol...

For subnanomolar libraries we use the protocol found in the supplemental section (Protocol 12) in the paper below. Its a fantastic protocol and I have used it for libraries with 100 pM (diluted to 4...
Forum: Core Facilities 12-10-2014, 06:09 AM
Replies: 22
Views: 18,874
Posted By acockburn
I like the epMotion 5075. I think its a great...

I like the epMotion 5075. I think its a great little robot, especially if you are new to programming these types of devices. I like that it checks to make sure the proper labware is being used and...
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