I think that the problem may be cause in library preparation step. Can i ask you some question?
What 's application did you perform? Short gun or Amplicon sequencing?
If you perform Amplicon sequencing, How long does your template?
Mixed read may be cause by the short fragment presented in your library. You should remove the short-fragment and checked by using bioanalyzer before perform a next run.
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yes i am....i have sent all my data and LOT numbers of all my kits and waiting for their reply
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I hope you are already talking with Roche tech support if you suspect the kits.Leave a comment:
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More mixed reads....help urgent
Hello,
i have been lately facing problems with my junior run. i have more mixed reads in my run which is like 50%. i performed 3 runs, cpb is 1 and all have enrichemnt above 20% (~35%).
In my last run i have reduced my cpb to half (0.5cpb) but it is still the same.
previously it used to work perefctly fine but recently all the problems are showing up. i use kapa qPCR for quantification.
onemore issue is during the bead enrichemnt process...washes with the enhancing buffer does not yield the white DNA pellets like it used to before...literally sometimes no pellet after 2 washes.... Maybe a problem with the kits?
i used the lot nos 93926520 and 93931420...... anyone encountering problems with these lot numbers.
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Amplicon sequencing is a targeted approach that allows researchers to investigate specific regions of the genome. This technique is routinely used in applications such as variant identification, clinical research, and infectious disease surveillance. The amplicon sequencing process begins by designing primers that flank the regions of interest. The DNA sequences are then amplified through PCR (typically multiplex PCR) to produce amplicons complementary to the targets. RNA targets...03-21-2023, 01:49 PM -
Targeted sequencing is an effective way to sequence and analyze specific genomic regions of interest. This method enables researchers to focus their efforts on their desired targets, as opposed to other methods like whole genome sequencing that involve the sequencing of total DNA. Utilizing targeted sequencing is an attractive option for many researchers because it is often faster, more cost-effective, and only generates applicable data. While there are many approaches...03-10-2023, 05:31 AM
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