SEQanswers

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



Similar Threads
Thread Thread Starter Forum Replies Last Post
Using Clean NGS from proteigene for RNA clean up of single cell Kaizen Sample Prep / Library Generation 0 07-31-2017 12:00 AM
Ampure bead clean up anna_m Sample Prep / Library Generation 7 06-20-2013 03:47 AM
96 well PCR clean up HMorrison Sample Prep / Library Generation 2 07-02-2012 06:32 PM
Clean room or not? denice_please Core Facilities 2 05-21-2012 01:45 AM
PCR Clean up - 96 well HGENETIC Sample Prep / Library Generation 0 07-26-2010 01:44 AM

Reply
 
Thread Tools
Old 02-18-2019, 02:58 AM   #1
Marjun Berbisá
Junior Member
 
Location: Faroe Islands

Join Date: Feb 2019
Posts: 1
Default are the samples clean enough for sequencing

Hello

Here are my questions:
I am collecting stool samples from patients and preparing v4 amplicon libraries for sequencing on the MiSeq using the goLay barcode strategy with primer set 515F/806R which expects a size around 380-400

1. How do you determine if the samples are clean enough for sequencing? I´m concerned about doing multiple rounds of Ampure bead clean-up and losing all my PCR product if it isn´t necessary. I have attached the Bioanalyzer traces for reference.
2. As a trial run, we are thinking about running 21 samples + positive and negative controls with 50% PhiX to be super conservative just to see that we get usable data. We plan on using the V2-300 cycle MiSeq kit and the new 3.1 software is getting installed this week. What is your advice on this? Do you have a step-by step instruction on how to set up the run using the Golay barcode method and these samples?
Attached Files
File Type: pdf 2100_DNA 1000_DNA 1000_DE13806351_2019-01-18_15-18-56.pdf (361.5 KB, 22 views)
Marjun Berbisá is offline   Reply With Quote
Reply

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 01:38 PM.


Powered by vBulletin® Version 3.8.9
Copyright ©2000 - 2019, vBulletin Solutions, Inc.
Single Sign On provided by vBSSO