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   Sample Submission Guidelines
   

Sanger Sequencing

 

Sanger Sequencing is a cost-effective method for determining the nucleotide sequence of DNA. GENEWIZ Sanger sequencing services are award-winning, providing high-quality results, industry-leading customer service and fast turnaround times at competitive prices. GENEWIZ from Azenta Life Sciences is the partner of choice for academic, pharmaceutical, GLP, biotechnology, and government institutions around the world.

We are well-equipped to handle DNA sequencing of all complexities including single tube, high-throughput, direct bacterial colony, glycerol stock, CLIA, or GLP DNA sequencing projects. Our mission is to eliminate bottlenecks, improve productivity, and advance your research progress.

GENEWIZ Sanger Sequencing Service

GENEWIZ accepts a variety of different templates to use as starting material for Sanger sequencing reactions.
Click on the service below to view the service options available for each starting material.

Sanger Sequencing for Purified Templates

GENEWIZ provides reliable, award-winning Sanger DNA Sequencing for purified templates through our Pre-Mixed, Pre-Defined, Custom and Difficult Template sequencing service options.

▼Pre-Mixed


  • For sequencing plasmids or purified PCR products, this service is the most economical choice.
  • You provide your template and primer mixed in the same tube according to our Sample Submission Guidelines.
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▼Pre-Defined


  • By choosing our Pre-Defined Service, you submit your template and primer in separate tubes, and a GENEWIZ scientist will mix them when your sample is received. An additional charge applies in most cases.
  • For sequencing plasmids or purified PCR products, submit your template and primer in separate tubes according to our Sample Submission Guidelines.
  • A GENEWIZ Universal Primer can be used in this service at no charge.
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▼Custom


  • Full-service option for sequencing of purified plasmids and purified PCR product samples with unknown concentration. For purified plasmids column-based PCR purifications, an GENEWIZ scientist will measure and adjust the concentration of your template to optimize the sequencing reaction.
  • Note that if you have enzymatically-purified PCR products, we recommend that you submit these samples under the Pre-Mixed or Pre-Defined service levels. An accurate OD cannot be obtained from an enzymatically-purified templates.
  • PCR Purification
    • Our enzymatic PCR clean-up technique removes excess primers and unincorporated dNTPs from PCR sequencing templates prior to the cycle sequencing reaction.
    • GENEWIZ uses a combination of enzymes to remove excess dNTPs and oligonucleotides from the PCR reaction prior to sequencing.
    • This approach generates a good-quality template that is ready to sequence, while avoiding common contamination problems from other, column-based purification techniques.
  • Submit template and primer separately according to our Sample Submission Guidelines.
  • A GENEWIZ Universal Primer can be used in this service.
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▼Difficult Template Sequencing


  • Specifically designed for challenging templates featuring GC-rich regions or intricate secondary structures like hairpins. With a success rate surpassing standard workflows by over double, researchers can now save at least a day in their process. Versatile and efficient, this protocol accommodates various DNA types without the need for additional templates, offering unparalleled convenience and reliability.

Template examples that may benefit from the alternative protocols include:

  • GC-rich: A template of > 60% GC content or high GC-content concentrated in a small region
  • Hairpin Structures: A sequence containing two inverted repeats, separated by at least three nucleotides
  • Repeats: Certain di- or tri-nucleotide repeats
  • Motifs causing band compressions: Mainly 5′-YGN1-2AR motifs, (Y: pyrimidine; R: purine; N: any base)
  • >Bacterial Artificial Chromosomes (BAC): Any construct larger than 50 kb.
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▼Primer Walking


Primer Walking Services

  • Utilizes multiple primers in a series to sequence entire plasmids or PCR products
  • Service options include plasmid sequence confirmation for already known sequences and plasmid sequence discovery for unknown sequences
  • For AAV vectors, we offer primer walking plus our new, proprietary AAV-ITR Sequencing
Plasmid Sequence Confirmation Service Plasmid Sequence Discovery Service
  • Confirm the sequence of your cloned products/genes of interest
  • Sequence your entire plasmid, including difficult-to-amplify regions
  • Examine failed amplifications and incorrect restriction digest patterns
  • Sequence full length AAV plasmids and hard to sequence ITR regions
  • Create a plasmid map for your undetermined plasmids
  • Confirm the sequence of your mystery constructs
  • Identify unknown regions of your plasmids
  • Sequence full length AAV plasmids and hard to sequence ITR regions

Clinical Sanger Sequencing

CLIA Sanger sequencing as the clinical “gold standard” from our state-of-the-art CLIA-certified and CAP-accredited laboratory while following Good Clinical Practices (GCP) guidelines.

▼CLIA Sanger Sequencing


  • Standalone sequencing service utilizes a CLIA-compliant workflow to support your clinical diagnostic assays, leading to advancements in personalized medicine and molecular diagnostics
  • Best utilized as the initial readout from a PCR assay or in conjunction with next generation sequencing, specifically to elucidate complex regions not approachable by other methodologies or to confirm variants uncovered by genome-wide techniques
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▼Regulatory Sanger Sequencing


Standalone Sanger sequencing services are also offered at the GLP-compliant level. For more information, please email regulatory@azenta.com

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Sanger Sequencing Features & Benefits

High Quality – DNA sequencing read lengths up to ~1000 bases. A typical read will provide 800 bases Phred20. Quality Score and Contiguous Read Length (CRL) provided in each read.
GENEWIZ Technical Support – Our highly-trained team of Sanger sequencing specialists proactively communicate updates, while providing technical expertise on projects big and small.
Convenient – Contact us to locate a drop box or pick-up service near you for free shipping.
Easy, Online Ordering – 24/7 web-based online ordering and data retrieval, with bar-coded order tracking make it easy to place your order and view data.
Best-in-Class Difficult Template Protocols – Our proprietary technologies allow for consisent, reliable data even on the most difficult templates (including hairpins and GC-rich DNA).
Free Vector Primers – a wide selection of GENEWIZ Universal Primers are available for most standard vectors. Also visit our Universal Primer Selection Tool to determine which Universal Primer binds to your vector of choice.

Sanger Sequencing Protocol

* Samples must arrive at the GENEWIZ from Azenta New Jersey laboratory before 10:00 am EST to qualify for Same Day service. Note that direct-sequencing templates are not available for our Same Day service.

Same Day

FOR RUSH PROJECTS: Data is reported by 7PM Eastern the same day we receive samples. A small additional charge applies. Please contact Technical Support for additional information.

Overnight Local Service

Overnight service for customers near a GENEWIZ laboratory. Laboratories are located in New York City, Boston, Washington D.C. Metro Area, Research Triangle Park, San Diego, Los Angeles Metro Area, San Francisco Bay Area, and Seattle. Typically samples are picked up in the afternoon by a GENEWIZ courier, and data are reported the next morning. Please contact us for details and eligibility.

Standard 24-Hour

Data reported within one business day after samples are received by GENEWIZ from Azenta.

Technical Resources

 

 

Sanger Sequencing Quick Tips Guide

Learn from the Sanger experts how you can get the most of your DNA Sequencing. Whether you’re troubleshooting a tough sequencing reaction, perfecting PCR visualization, or learning to clean up and refine your PCR products, these resources are designed to help you get the best results.
• Volume 1: Producing Robust, Single-band PCR Product
• Volume 2: PCR Clean-up of Single-band PCR
• Volume 3: Salvaging Nonspecific PCR Products
• Volume 4: PCR Visualization by Gel Electrophoresis
• Volume 5: Troubleshooting a Bad Sequencing Reaction from a PCR Template

Blog | Troubleshooting DNA Templates with Sanger Sequencing

Sanger sequencing is highly effective for testing small targeted genomic regions and for validating results from NGS. However, as with all sequencing technologies, there are limitations and needs for troubleshooting. Read on to learn about the three basic troubleshooting steps we recommend to start with.

Blog | Ensure Complete Plasmid Validation on Your First Attempt with Oxford Nanopore Sequencing

Explore the cutting-edge realm of genomic research with Oxford Nanopore Sequencing (ONT), and how this innovative Next Generation Sequencing technology offers real-time, long-read capabilities, revolutionizing our approach to decoding genetic information.

Webinar | From Base Pairs to Breakthroughs: Understanding Sanger and Oxford Nanopore Sequencing

While Sanger sequencing remains the gold standard for accurate DNA sequencing, advancements in sequencing technology, such as Oxford Nanopore Technology (ONT), have provided a cost-effective, long-read alternative for analysis. In this workshop, delve into the considerations of each technique and discover how to interpret their resulting sequencing data effectively so you can capture the benefits of both approaches for your specific needs.

Blog | Analyzing Sanger Sequencing Data

The output for Sanger sequencing is typically a chromatogram, also known as a trace or ab1 file, and a text-based sequence file. Although the latter may seem to hold all the relevant information—after all, the point of sequencing is to get a sequence—the former can’t be ignored. Here, we provide a guide to understanding Sanger sequencing data.