Go Back   SEQanswers > Sequencing Technologies/Companies > 454 Pyrosequencing

Similar Threads
Thread Thread Starter Forum Replies Last Post
best primers for 16S amplicon sequencing dca 454 Pyrosequencing 9 09-11-2012 09:49 AM
amplicon sequencing with Lib-L (Junior) audioPL 454 Pyrosequencing 9 11-27-2011 04:53 PM
PubMed: Barcoded primers used in multiplex amplicon pyrosequencing bias amplification Newsbot! Literature Watch 0 09-06-2011 02:00 AM
GS Junior enrichment Bucky 454 Pyrosequencing 1 10-19-2010 12:15 AM
Roche Junior amplicon adapters jffkm General 0 02-11-2010 02:22 PM

Thread Tools
Old 01-12-2011, 10:33 PM   #1
Junior Member
Location: Aachen

Join Date: Jan 2011
Posts: 3
Default GS Junior - amplicon primers

Hi all,

we just got our GS Junior and I am currently designing primers for amplicon sequencing. Does anyone have some experience with it? For example which amplicon size is acceptable for biderectional sequencing? Is there a cut of in primer length an GC content? What is the maximum acceptable size difference for amplicons in emPCR not affecting the coverage? I know that Roche gives some parameters but im really interested to know what will work in real life.

Thank You all!
mikebravo is offline   Reply With Quote
Old 02-28-2011, 06:32 AM   #2
Junior Member
Location: germany

Join Date: Feb 2011
Posts: 1

Hi mikebravo,


if you wish to do bidirectional sequencing an amplicon length of about 400 bp is acceptable, i think. In our training course we were advised to minimize the size differences to a maximal range of 100 bp. We use amplicons with a length ranging from 370 to 470 bp. In general all requirements that are essential for all-day primer design have to be taken into consideration when designing your NGS-Primer. The length of my primers range between 50 and 58 bp (with MID, Key and A or B). I wish you good luck for your experiments with your GS Junior.

Greetings from Berlin
junioshi is offline   Reply With Quote

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 02:05 PM.

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