This kit has not been tested in-house with PFA-fixed samples. However, academic literature indicates successful use of this kit with fixed samples.
WGA4
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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10 REACTIONS
$284.00
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$1,120.00
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$9,980.00
$284.00
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About This Item
UNSPSC Code:
12352200
EC Number:
254-457-8
NACRES:
NA.55
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dilution
(WGA)
input
purified DNA
shipped in
wet ice
technique(s)
whole genome amplification: suitable
Quality Level
storage temp.
−20°C
Related Categories
1 of 4
This Item | |||
|---|---|---|---|
| technique(s) whole genome amplification: suitable | technique(s) whole genome amplification: suitable | technique(s) whole genome amplification: suitable | technique(s) whole genome amplification: suitable |
| shipped in wet ice | shipped in wet ice | shipped in wet ice | shipped in wet ice |
| storage temp. −20°C | storage temp. −20°C | storage temp. −20°C | storage temp. −20°C |
| Quality Level 200 | Quality Level 200 | Quality Level 200 | Quality Level 200 |
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
- in whole genome amplification (WGA)
- to amplify the microdissected DNAs
- for the amplification of circulating tumor cell genomic DNA from metastatic castration-resistant prostate cancer cells[1]
- for the detection of copy number variations (CNV) by single-cell low-coverage whole-genome sequencing (SLWGS) method[2]
- 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.
related product
Product No.
Description
Pricing
signalword
Danger
hcodes
pcodes
Hazard Classifications
Resp. Sens. 1
Storage Class
10 - Combustible liquids
flash_point_f
Not applicable
flash_point_c
Not applicable
wgk
WGK 3
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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
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
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
Timour Baslan et al.
eLife, 9 (2020-05-14)
Copy number alterations (CNAs) play an important role in molding the genomes of breast cancers and have been shown to be clinically useful for prognostic and therapeutic purposes. However, our knowledge of intra-tumoral genetic heterogeneity of this important class of
Articles
Whole genome amplification overcomes restrictions for single-cell genomic analyses with non-specific amplification.
Protocols
Whole genome amplification (WGA) of plasma and serum DNA presents a unique challenge due to the small amount of nucleic acid in such samples.
GenomePlex® is a Whole Genome Amplification (WGA) method that allows the researcher to generate a representative amplification of genomic DNA
This protocol provides a simple and convenient method to isolate, amplify, and purify genomic DNA from buccal swabs. Buccal swabs are a convenient method of acquiring a DNA sample.
Related Content
Our commitment to the Life Sciences continues through significant advances in PCR technology. Improvements to the key parameters of yield, length, specificity, and fidelity, as well as new applications for nucleic acid amplification form the basis of our research and development focus. We are dedicated to providing you with the newest technology as the field of Nucleic Acid Amplification evolves, making Sigma your preferred supplier for PCR products.
Instructions
Chromosome Analysis of Single Cells by High Resolution Comparative Genomic Hybridization after Whole Genome Amplification Using a Random Fragmentation Approach
Single Cell Aneuploidy Analysis by High Resolution Comparative Genomic Hybridization (CGH) after Whole Genome Amplification reveals Maternal Cell Contamination in the Original Products of Conception Cultures.
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Is this kit compatible with PFA-fixed samples or should I first reverse crosslinked DNA and then use a broad-spectrum DNA repair enzyme cocktail before doing WGA?
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Buongiorno, il kit GenomePlex® Single Cell WGA4 per amplificazione di DNA da singola cellula con quali metodiche di preparazione di librerie è compatibile? Vi ringrazio
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The GenomePlex® Single Cell WGA4 Single Cell DNA Amplification Kits can be sequenced using any library preparation kit that includes end repair (Blunting + A tailing + 5' phosphorylation) followed by adapter ligation.
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I want the amplified product to be NGS sequenced. What is the error rate during amplification and the coverge rate of the genome?
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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).
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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-
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.
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What A260/A280 ratios are expected for a successful WGA 4 product using the WGA4 kit?
1 answer-
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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Can Product WGA4, GenomePlex® Single Cell Whole Genome Amplification Kit, be used with purified DNA?
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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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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-
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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What is the Department of Transportation shipping information for this product?
1 answer-
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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What kinds of cells can be used as starting materials with Product WGA4, GenomePlex® Single Cell Whole Genome Amplification Kit?
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The cell lysis method involves treatment with proteinase K and heat. Most cells will be lysed under these conditions.
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