GenomePlex® Single Cell Whole Genome Amplification Kit
SIGMA/WGA4 - Amplify genome of a single cell
Synonym: Single cell whole genome amplification; Whole genome amplification
EC Number: 254-457-8
Product Type: Chemical
U937 Leukemia Single Cells were amplified using the GenomePlex Single Cell WGA procedure. Ten nanograms of amplified DNA was then utilized in a MGB Eclipse SNP quantitative PCR reaction on the ABI 7700. Dissociation curves allow allele determination based upon the melting temperature (Tm). FAM and TET fluorescent signals distinguish homozygous wild and heterozygous mutant types. The WGA amplified single cell DNA was tested for the NAT1-01 SNP as well as MTHFR01 and IGF1R-03 (data not shown). Unamplified control DNA was evaluated with each group to serve as a positive control. The data indicates that all amplified single cells were the same allele for each SNP tested and were, therefore, unchanged by WGA.
DNA from normal and tumorigenic human kidney cells were amplified using the GenomePlex Single Cell WGA Kit. Amplified material was hybridized to metaphase BAC arrays to determine chromosomal karyotype. Data that falls outside of the red line indicates chromsomal loss, while data that continues past the green line suggests chromosomal amplification. As expected the control sample demonstrated a balanced chromosomal copy number. Chromosome 3 for the amplified kidney tumor single cell displayed under representation as depicted by the red bar. These results match previous microarray work using an amplification method that took three days. GenomePlex technology accurately amplifies genomic material down to single cell resolution. Data is courtesy of Dr. Michael Speicher from Institute Human Genetics, TU Munich.
Kit Image
| Quality Level | 200  |
| shipped in | wet ice |
| storage temp. | −20°C |
| technique(s) | whole genome amplification: suitable |
| 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 • for the detection of copy number variations (CNV) by single-cell low-coverage whole-genome sequencing (SLWGS) method • 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 |
| 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. |
| Legal Information: | GenomePlex is a registered trademark of Takara Bio USA, Inc. |
| Legal Information: | TaqMan is a registered trademark of Roche Molecular Systems, Inc. |
| 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. |
| Other Notes: | The sequences of the universal primers provided in this kit are considered proprietary. |
| Symbol |  GHS08 |
| Signal word | Danger |
| Hazard statements | H334 |
| Precautionary statements | P261 - P284 - P501 |
| Hazard Codes | Xn |
| Risk Statements | 36/37/38-42 |
| Safety Statements | 22-24-26-36/37 |
| RIDADR | NONH for all modes of transport |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| UNSPSC | 12352200 |
| Components | 10x Single Cell Lysis & Fragmentation Buffer; Proteinase K from Tritirachium album, buffered aqueous glycerol solution, for molecular biology, ≥800 units/mL; Water, Nuclease-Free Water, for Molecular Biology |