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WGA4

Sigma-Aldrich

GenomePlex® Single Cell Whole Genome Amplification Kit

Amplify genome of a single cell

Synonym(s):

Single cell whole genome amplification, Whole genome amplification

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About This Item

EC Number:
254-457-8
UNSPSC Code:
12352200
NACRES:
NA.55
Pricing and availability is not currently available.

Quality Level

200

technique(s)

whole genome amplification: suitable

shipped in

wet ice

storage temp.

−20°C

Related Categories

General description

GenomePlex® Single Cell Whole Genome Amplification Kit utilizes a proprietary technology based on random fragmentation of genomic DNA and conversion of the resulting small fragments to PCR-amplifiable library molecules flanked by universal priming sites. WGA is achieved by PCR amplification of the library molecules using universal oligonucleotide primers. This kit is designed to amplify the genome of a single cell. This rapid and straightforward method provides millionfold amplification yielding microgram quantities of genomic DNA from a single cell. Traditional single-cell whole genome amplification methods yield insufficient quantities with significantly biased representation. The kit includes all the reagents necessary for cell lysis and successive whole genome amplification. Single cells can be isolated by fluorescence-activated cell sorting (FACS), laser capture microdissection (LCM), dilution, or any other applicable method.

Application

GenomePlex® Single Cell Whole Genome Amplification Kit has been used:

  • to amplify the isolated DNA[1]
  • in whole genome amplification (WGA)[2]
  • to amplify the microdissected DNAs[3]
  • for the amplification of circulating tumor cell genomic DNA from metastatic castration-resistant prostate cancer cells[4]
  • for the detection of copy number variations (CNV) by single-cell low-coverage whole-genome sequencing (SLWGS) method[5]
  • Gel electrophoresis
  • qPCR
  • comparative genomic hybridization (CGH) microarray
  • Short tandem repeats (STR) analysis
  • single nucleotide polymorphism (SNP) analysis

Features and Benefits

  • Highly yield and accuracy of DNA amplification within four hours
  • Amplification of DNA from any source such as cancer cells, epithelial cells, lymphocytes, fibroblast amniotic cells, polycarbonate fixed cells, and plant cells
  • A complete representation of the entire genome with minimal allele bias
  • WGA DNA polymerase is suitable for use with downstream applications including gel electrophoresis, qPCR, comparative genomic hybridization (CGH) microarray, short term repeat (STR) analysis, and single nucleotide polymorphism (SNP) analysis, TaqMan® assays, and microsatellite analysis

Other Notes

The sequences of the universal primers provided in this kit are considered proprietary.

Legal Information

Use of this product is covered by one or more of the following US patents and corresponding patent claims outside the US: 5,789,224, 5,618,711, 6,127,155 and claims outside the US corresponding to expired US Patent No. 5,079,352. The purchase of this product includes a limited, non-transferable immunity from suit under the foregoing patent claims for using only this amount of product for the purchaser′s own internal research. No right under any other patent claim, no right to perform any patented method, and no right to perform commercial services of any kind, including without limitation reporting the results of purchaser′s activities for a fee or other commercial consideration, is conveyed expressly, by implication, or by estoppel. This product is for research use only. Diagnostic uses under Roche patents require a separate license from Roche. Further information on purchasing licenses may be obtained by contacting the Director of Licensing, Applied Biosystems, 850 Lincoln Centre Drive, Foster City, California 94404, USA.
GenomePlex is a registered trademark of Takara Bio USA, Inc.
TaqMan is a registered trademark of Roche Molecular Systems, Inc.

Kit Components Also Available Separately

Product No.
Description
SDS
  • L104310x Single Cell Lysis & Fragmentation BufferSDS
  • P4850Proteinase K from Tritirachium album, buffered aqueous glycerol solution, for molecular biology, ≥800 units/mLSDS
  • W4502Water, Nuclease-Free Water, for Molecular BiologySDS

Pictograms

Health hazard
GHS08

Signal Word

Danger

Hazard Statements

H334

Precautionary Statements

P261 - P284 - P501

Hazard Classifications

Resp. Sens. 1

Storage Class Code

10 - Combustible liquids

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Choose from one of the most recent versions:

Certificates of Analysis (COA)

Lot/Batch Number
0000331177
0000331177
SLCR4442
SLCR4442
SLCP0781

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Resolving tumor heterogeneity: genes involved in chordoma cell development identified by low-template analysis of morphologically distinct cells
El-Heliebi A, et al.
PLoS ONE, 9(2), e87663-e87663 (2014)
Heike Fiegler et al.
Nucleic acids research, 35(3), e15-e15 (2006-12-21)
Heterogeneity in the genome copy number of tissues is of particular importance in solid tumor biology. Furthermore, many clinical applications such as pre-implantation and non-invasive prenatal diagnosis would benefit from the ability to characterize individual single cells. As the amount
Nathan R Treff et al.
Molecular human reproduction, 16(8), 583-589 (2010-05-21)
Many studies estimate that chromosomal mosaicism within the cleavage-stage human embryo is high. However, comparison of two unique methods of aneuploidy screening of blastomeres within the same embryo has not been conducted and may indicate whether mosaicism has been overestimated
Nathan R Treff et al.
Fertility and sterility, 94(6), 2017-2021 (2010-03-02)
To develop and validate a whole genome amplification and single nucleotide polymorphism (SNP) microarray protocol for accurate single cell 24 chromosome aneuploidy screening. Prospective, randomized, and blinded study. Academic reproductive medicine center. Multiple euploid and aneuploid cell lines were obtained
Anna A Torgasheva et al.
Proceedings of the National Academy of Sciences of the United States of America, 116(24), 11845-11850 (2019-05-01)
An unusual supernumerary chromosome has been reported for two related avian species, the zebra and Bengalese finches. This large, germline-restricted chromosome (GRC) is eliminated from somatic cells and spermatids and transmitted via oocytes only. Its origin, distribution among avian lineages
View All Related Papers

Articles

Whole Genome Amplification for Single Cell Biology

Whole genome amplification overcomes restrictions for single-cell genomic analyses with non-specific amplification.

Whole Genome Amplification for Single Cell Biology

Whole genome amplification overcomes restrictions for single-cell genomic analyses with non-specific amplification.

Whole Genome Amplification for Single Cell Biology

Whole genome amplification overcomes restrictions for single-cell genomic analyses with non-specific amplification.

Whole Genome Amplification for Single Cell Biology

Whole genome amplification overcomes restrictions for single-cell genomic analyses with non-specific amplification.

Protocols

GenomePlex® Single Cell Whole Genome Amplification Kit (WGA4) - Protocol

GenomePlex® is a Whole Genome Amplification (WGA) method that allows the researcher to generate a representative amplification of genomic DNA

Whole Genome Amplification from Serum or Plasma Protocol

Whole genome amplification (WGA) of plasma and serum DNA presents a unique challenge due to the small amount of nucleic acid in such samples.

Whole Genome Amplification from Serum or Plasma Protocol

Whole genome amplification (WGA) of plasma and serum DNA presents a unique challenge due to the small amount of nucleic acid in such samples.

Whole Genome Amplification from Serum or Plasma Protocol

Whole genome amplification (WGA) of plasma and serum DNA presents a unique challenge due to the small amount of nucleic acid in such samples.

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Questions

1–7 of 7 Questions  

  1. I want the amplified product to be NGS sequenced. What is the error rate during amplification and the coverge rate of the genome?

    1 answer

    1. The error rate during amplification has not been determined, but without proofreading, the whole genome amplification polymerase error rate is expected to be similar to native Taq polymerase, typically in the range of approximately 1 × 10^5 to 2 × 10^4. Publications specifically quantifying coverage can be found in early work with WGA4, which reported an overall measured coverage of 78% for aneuploidy screening (doi.org/10.1016/j.fertnstert.2007.07.789). Later work using WGA4 to measure gene copy number variation (https://doi.org/10.1038/srep11415) showed coverage varied by GC content; in areas with a GC composition of 30-60%, the coverage varied approximately 60-120% (rat genomic DNA has 41.9% GC). Several papers have been published on obtaining optimal results and accurate analysis from the kit, with the most relevant being (https://doi.org/10.1038/nprot.2012.039).

      Helpful?

  2. Is there an estimate for the ratio of ssDNA to dsDNA, like roughly 3 times more ssDNA than dsDNA? Also, does the advertised 5-10 ug of DNA for WGA4 correspond to total DNA or just dsDNA?

    1 answer

    1. The ratio of ssDNA to dsDNA has not been determined. However, the 5-10 ug of DNA advertised for WGA4 corresponds to total DNA, not just dsDNA.

      Helpful?

  3. What A260/A280 ratios are expected for a successful WGA 4 product using the WGA4 kit?

    1 answer

    1. The A260/280 has not been measured for any amplification product; however, given that starting material is infinitesimal quantities and adding very little protein, the expectation is for A260/280 to resemble that of pure DNA, especially after purification using the GenElute PCR Cleanup Kit (Catalog Number NA1020).

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  4. Can Product WGA4, GenomePlex® Single Cell Whole Genome Amplification Kit, be used with purified DNA?

    1 answer

    1. Yes. The kit is designed to amplify the small amount of DNA that is present in a single cell (3-5 pg). It will also work with small amounts of purified DNA; we recommend less than 100 pg.

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  5. Can the amplified Omniplex products resulting from the use of Product WGA4, GenomePlex® Single Cell Whole Genome Amplification Kit, be used for TA cloning?

    1 answer

    1. Yes.  Please ensure that a final extension of 7-30 minutes at 72°C is included after amplification if performing subsequent TA cloning reactions.

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  6. What is the Department of Transportation shipping information for this product?

    1 answer

    1. Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.

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  7. What kinds of cells can be used as starting materials with Product WGA4, GenomePlex® Single Cell Whole Genome Amplification Kit?

    1 answer

    1. The cell lysis method involves treatment with proteinase K and heat. Most cells will be lysed under these conditions.

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