SINGLE CELL WHOLE GENOME AMPLIFICATION

DOPlify® WGA V2 Kit & Technology

The DOPlify® WGA V2 kit performs fast whole genome amplification on single cells or limited template DNA samples, allowing for the cell chromosome copy number status to be determined. The DOPlify® WGA V2 kit offers the advantage of full mitochondrial DNA coverage and the ability to add custom primers to specially amplify regions of interest. Products of the DOPlify® WGA V2 kit can be used for a wide range of downstream processes, workflows, and NGS applications. The technology captures exclusive know-how for the lysis and whole genome amplification, with reagents provided in an easy-to-use kit format and less than 3 hour protocol time.

  • NGS compatible advanced DOP-PCR
  • Fast whole genome amplification with protocol time < 3 hours
  • Reduced contamination risk with only 2 tube openings required and 3 reagents to add
  • Can be fully automated with the JANUS® G3 liquid handler
  • No amplification of No Template Control
    • 2.5 μg of DNA from one cell

  • 4321-0020

    50 RXNS
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For research use only. Not for use in diagnostic procedures.

Why Use DOPlify® Technology?

The DOPlify® WGA V2 kit is a whole genome amplification kit designed to PCR amplify the genome from single cells and limited DNA templates.

The DOPlify® kit is unique in that it uses advanced DOP-PCR, which;

  • uses the high processivity and fidelity of new generation polymerases that are NGS application grade
  • uses ready-mixes to minimize the pipetting steps and requiring only two sample tube openings to reduce the risk of contamination
  • provides an amplification system that is flexible enough to concurrently whole genome amplify a sample while specifically amplifying genes of interest, reducing the risk of allele drop-out.
DOPlify v2 Workflow

DOPLIFY® WORKFLOW

Protocol Step Explanation Time
Cell Lysis A gentle but effective enzyme-based lysis procedure ensures robust cell lysis and a readily accessible DNA template for whole genome amplification 15 mins
Whole Genome Amplification Whole genome amplification is performed using advanced DOP-PCR, which has been optimized for limited DNA input. DOP-PCR uses degenerate primers to initiate DNA amplification, binding across a broad range of different sequences scattered genome wide 2.5 hrs
Quality Control Following amplification, the analysis of products by agarose gel electrophoresis or microfluidic electrophoresis is recommended to ensure that cell amplification has been successful 30 mins

Can DOPlify® WGA V2 be Automated?

The whole genome amplification of single or small numbers of cells and downstream processes such as library preparation for NGS can be fully automated with the JANUS® G3 liquid handler.

Whole genome amplification (WGA) of single cells in tubes has traditionally been performed by hand, which can lead to multiple potential issues including:

  • Errant volume pipetting
  • Technologist fatigue
  • Wasted time
  • Sample contamination

Using the JANUS® G3 liquid handler with preprogrammed, easy-to-use automated WGA protocols your lab can:

  • Reduce human errors and contamination risk
  • Gain time for your staff to perform more important tasks
  • Increase your lab’s throughput with walkaway automation
  • Ensure sample process traceability with barcode scanning and sample tracking

PerkinElmer’s new custom JANUS® G3 liquid handler format is capable of performing WGA using the DOPlify® platform or other WGA technologies with a throughput of 48 – 96* samples per run.

* Higher throughput options are available

KIT CONTENTS

  • Cell Lysis Mix
  • WGA Ready-mix
  • WGA Primer

REQUIRED MATERIALS NOT PROVIDED

  • 2 or 0.5 mL PCR tubes
  • Single blastomeres, polar bodies, trophoblastic cells, amniocytes, lymphocytes, or cultured cells
  • Pipettes
  • PCR thermocycler with programmable ramp rate

Publications that Cite Using the DOPlify® Kit for Whole Genome Amplification

Deleye, L., Tilleman, L., Plaetsen, A. V., Cornelis, S., Deforce, D., & Nieuwerburgh, F. V. (2017). Performance of four modern whole genome amplification methods for copy number variant detection in single cells. Scientific Reports, 7(1). doi:10.1038/s41598-017-03711-y.

Deleye, L., Plaetsen, A. V., Weymaere, J., Deforce, D., & Nieuwerburgh, F. V. (2018). Short Tandem Repeat analysis after Whole Genome Amplification of single B-lymphoblastoid cells. Scientific Reports,8(1). doi:10.1038/s41598-018-19509-5.

Mai, A. D., Harton, G. L., Quang, V. N., Van, H. N., Thi, N. H., Thuy, N. P., . . . Quoc, Q. T. (2020). Development and clinical application of a preimplantation genetic testing for monogenic disease (PGT-M) for beta thalassemia in Vietnam. Journal of Assisted Reproduction and Genetics. doi:10.1007/s10815-020-02006-y.

Plaetsen, A. V., Deleye, L., Cornelis, S., Tilleman, L., Nieuwerburgh, F. V., & Deforce, D. (2017). STR profiling and Copy Number Variation analysis on single, preserved cells using current Whole Genome Amplification methods. Scientific Reports, 7(1). doi:10.1038/s41598-017-17525-5.

The DOPlify® WGA V2 Kit contains reagents for amplifying 50 samples. Upon receipt, store the kit at -20°C in a constant temperature freezer (not frost free).

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