Sanger Sequencing

Sample Submission Guidelines

DNA Sequencing Sample Submission Guidelines

For the best Sanger sequencing results, Azenta Life Sciences, formerly GENEWIZ, strongly recommends you follow our Sample Submission Guidelines as closely as possible. If you have any questions, our Technical Support team is here to help!

(EU) Email: sanger.europe@azenta.com | Phone +49 (0)341 520 122-41

(UK) Email: genomics.service.uk@azenta.com | Phone +44 (0) 1279 873837

Sample Preparation

Follow our barcode labeling instructions to avoid delays in sample processing and data delivery:

For orders with templates and primers in tubes, please use 1.5 ml flip cap reaction tubes (no screw caps). Label your tubes with barcodes according to your selected sample type to facilitate sample alignment and automated processing. Please note we do not accept samples labeled with fluorescent or other dyes.

Dilute your custom sequencing primer to 10 or 5 µM (pmol/µl) according to the service type (Pre-Mixed, Pre Defined) using water. Please refer to the table below for minimum volume for each sequencing reaction.

For the amount of template required in each of our DNA Sequencing Services (Pre-Mixed, Pre-Defined), please refer to the tables below. Please make dilutions in water or Tris. For best results, do not use Tris-EDTA (TE) because EDTA will inhibit the sequencing reaction.

Pre-Mixed Tube or Plate/Sanger-EZ

For the Pre-Mixed Service, prepare purified template (DNA) and primer in a total volume of 10µl in the same tube according to the table below.

Template				Primer		Template + Your Primer
DNA Type	Submitted in	Concentration	Volume	Concentration	Volume*	Pre-Mixed Total Volume
Plasmids purified	1.5 ml reaction tube	30-100 ng / µl	5 µl	5 pmol/ µl (µM)	5 µl	10 µl
PCR fragments purified		10-50 ng / µl
Plasmid purified	96-well plate	30-100 ng / µl	5 µl	5 pmol/ µl (µM)	5 µl	10 µl
PCR fragments purified		10-50 ng / µl

Video | Pre-Mixed Tube Ordering Guide

This Sanger ordering guide explains how to place a Pre-Mixed Tube sequencing order via your Azenta account and label samples correctly.

Pre-Defined Tube

For Pre-Defined Services, provide template (20 µl) and your primer (20 µl) in separate tubes for up to eight reactions according to the table below.

Template				Primer
DNA Type	Submitted in	Concentration	Volume	Submitted in	Concentration	Volume
Plasmid purified	1.5 ml reaction tube	30-100 ng / µl	20 µl	1.5 ml reaction tubes	10 pmol/µl (µM)	20 µl
PCR fragments purified/ unpurified		10-50 ng / µl	20 µl

Video | Pre-Defined Tube Ordering Guide

This Sanger ordering guide explains how to place a Pre-Defined Tube sequencing order via your Azenta account, order primers and label samples correctly.

Pre-Defined Vario Plate

For Pre-Defined Sanger Sequencing service in 96-well plates, provide 20 µl template per well and for primer provide separately 100 µl per plate.

Template				Primer
DNA Type	Submitted in	Concentration	Volume	Submitted in	Concentration	Your Primer Volume Per Plate
Plasmid (purified)	96-well plate	30-100 ng/µl	20 µl	1.5 ml reaction tubes	10 pmol/µl (µM)	20 µl - for up to 8 reactions 100 µl - for more than 8 reactions
PCR product (purified / unpurified)		10-50 ng / µl	20 µl
Low concentration plasmid (EU)		10 ng / µl	20 µl

Video | Pre-Defined Vario Plate Ordering Guide

This Sanger ordering guide explains how to place a Vario Plate sequencing order via your Azenta account, order primers and label samples correctly.

Pre-Defined Plate

For Pre-Defined Sanger Sequencing service in 96-well plates, provide 20 µl template per well and for primer provide separately 100 µl per plate.

Template				Primer
DNA Type	Submitted in	Concentration	Volume	Submitted in	Concentration	Your Primer Volume Per Plate
Plasmids (purified)	96-well plate	30-100 ng/µl	20 µl	1.5 ml reaction tube	10 pmol/µl (µM)	100 µl - for more than 8 reactions
PCR product (purified / unpurified)		10-50 ng/µl	20 µl
Low concentration plasmid (EU)		10 ng/µl	20 µl

Pre-Mixed Vario Plate

For the Pre-Mixed Service, prepare purified template (DNA) and primer in a total volume of 10µl in the same tube according to the table below.

Template				Primer		Template + Your Primer
DNA Type	Submitted in	Concentration	Volume	Concentration	Volume*	Pre-Mixed Total Volume
Plasmid purified	96-well plate	30-100 ng/µl	5 µl	5 pmol/µl (µM)	5 µl	10 µl
PCR fragments Purified/ Unpurified		10-50 ng / µl

GENEWIZ SANGER Sequencing Logistic Options

GENEWIZ offers convenient logistic options for sample pick up or shipment to our European sequencing labs. Learn more about logistic options available.

Sanger Sequencing

Sample Preparation
Europe

Bacteria and New AAV-ITR
Europe

Primer Walking
Europe

Discover the Sequence for Success with Sanger-EZ

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DNA Sequencing Sample Submission Guidelines

Sample Preparation