PCR Plus Sequencing CRISPR Analysis Package

 

Confirming genome edits with Sanger sequencing is an important step in determining the efficiency of CRISPR-mediated genome editing. Our CRISPR Sanger sequencing service is designed to verify the successful edit of your region of interest, starting with cells or genomic DNA.

   



Troubleshooting DNA Templates
with Sanger Sequencing



 

 

CRISPR MUTATION ANALYSIS

Our Mutation Analysis service includes PCR optimization, amplicon production and purification, and Sanger sequencing for your CRISPR regions of interest. Whether you are confirming that a mutation has occurred or precisely determining the genotype and specific allelic event, leveraging our expertise ensures that your genome editing functions efficiently.

For CRISPR experiments, our experienced scientists can interrogate your specified on-target regions of interest. Azenta can also confirm off-target effects of the RNA-guided Cas9 nuclease1. Simply identify your target regions and let our Sanger experts handle the rest.

1 Nucleic Acids Research (2014) doi: 10.1093/nar/gku402. CRISPR/Cas9 systems have off-target activity with insertions or deletions between target DNA and guide RNA sequences. Lin, Y, et. Al.  

GENEWIZ PCR PLUS SEQUENCING CRISPR ANALYSIS SERVICES

Extraction of genomic DNA from single-isolate cell samples.

Amplicon selection and primer design for on- and off-target regions.
Assay development and optimization.
PCR amplification of genomic DNA.
High-throughput PCR purification and bi-directional DNA sequencing.
Automated and manual sequencing results analysis.
Report identifying nucleotide-level mutations.
Translated amino acid calls available upon request.

FEATURES & BENEFITS

Reliable Assay Development: Our highly-trained scientists develop PCR assays targeted to your region of interest.
Flexible Starting Materials: GENEWIZ can start with purified genomic DNA templates or extract it from supplied sources.

High-Throughput Processing: Target regions are amplified using optimized assays; amplicons are purified and sequenced by capillary electrophoresis in our best-in-class facility.

Accurate Data Analysis: Mutations are identified with both automated and manual analysis to ensure accuracy.

NGS, PCR, or Sanger Sequencing:
An Assay Selection Guide

Azenta vs. In-house Turnaround Time Comparison of PCR Sequencing CRISPR Analysis

AZENTA TURNAROUND: As fast as 5 DAYS

GENEWIZ TURNAROUND: As fast as 5 DAYS

IN-HOUSE TURNAROUND: 2-4 WEEKS

inhouse-turnaround-2-4-weeks

DELIVERABLES

  • – Amplicon selection and primer design to target regions of interest
  • – Assay development and optimization
  • – PCR amplification of genomic DNA using optimized conditions
  • – High-throughput PCR purification and bi-directional DNA sequencing per amplicon
  • – Automated and manual sequencing results analysis
  • – Report identifying mutations

HOW TO ORDER

*Samples must arrive at the 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.

Email | Phone (1-877-GENEWIZ (436-3949), Ext. 3350)