Go Back   SEQanswers > Applications Forums > Sample Prep / Library Generation

Similar Threads
Thread Thread Starter Forum Replies Last Post
Quantifying copies of a single locus robinweide Bioinformatics 0 10-17-2014 02:28 PM
The conversion of multiple copies seqs to single-copy lcy Bioinformatics 0 05-16-2013 09:37 PM
phrap/consed - multiple copies during assembly pag Bioinformatics 12 05-31-2012 10:26 AM
extract homeologuous copies from polyploids assemblies babine Bioinformatics 0 03-13-2012 04:43 AM

Thread Tools
Old 01-10-2015, 09:38 AM   #1
Junior Member
Location: Leuven, Belgium

Join Date: Apr 2012
Posts: 8
Default Diploid genomic copies actually sequenced

Hi all, I would like to know if there is a way to calculate how many different genomes you sequence when you prepare amplicons from human gDNA. The goal is to do a power-calculation to understand how much we need to sequence a sample to identify mosaic variants with low mutant allelic frequencies.

In a pilot experiment, we have prepared amplicons from 25 ng of template gDNA. Based on an average human diploid cell DNA content of 6.5 pg, this corresponds to ~3,846 copies of diploid genomes. To the resulting amplicon was then added P5 and P7 by PCR and sequenced on a MiSeq. Will my sequencing library have covered all the genomic copies used as template? Is there a way to calculate how many I have covered? Is there a library preparation step to control this parameter?

Thank you very much!
charlescoldroom is offline   Reply With Quote

amplicon, genomic copies, library

Thread Tools

Posting Rules
You may not post new threads
You may not post replies
You may not post attachments
You may not edit your posts

BB code is On
Smilies are On
[IMG] code is On
HTML code is Off

All times are GMT -8. The time now is 07:59 PM.

Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2020, vBulletin Solutions, Inc.
Single Sign On provided by vBSSO