Hi!
I encountered two problems while preparing Illumina's Trusight One library using their guide. I'm following the protocol carefully (except from adjusting the protocol to tubes instead of plates) and started from a total of 50ng of high quality human genomic DNA.
1. I'm getting low concentration (about half of the lower threshold) of the enriched library. The enriched library is prepared on a pool of 12-plex pre-enriched libraries, and the pre-enriched libraries are in good concentration (60-90 ng/ul each). I already repeated twice and got exactly the same results.
2. The library size is big. In the first time the average size (as measured by BioAnalyzer) was 600 bp, and in the second in was 520 bp. I'm measuring the input as accurately as possible DNA in triplicates using Qubit.
Need your advices, ideas and anything you can think that can help me improve this.
Thanks!
Merav
I encountered two problems while preparing Illumina's Trusight One library using their guide. I'm following the protocol carefully (except from adjusting the protocol to tubes instead of plates) and started from a total of 50ng of high quality human genomic DNA.
1. I'm getting low concentration (about half of the lower threshold) of the enriched library. The enriched library is prepared on a pool of 12-plex pre-enriched libraries, and the pre-enriched libraries are in good concentration (60-90 ng/ul each). I already repeated twice and got exactly the same results.
2. The library size is big. In the first time the average size (as measured by BioAnalyzer) was 600 bp, and in the second in was 520 bp. I'm measuring the input as accurately as possible DNA in triplicates using Qubit.
Need your advices, ideas and anything you can think that can help me improve this.
Thanks!
Merav