SeqPlex DNA Amplification Kit
SIGMA/SEQXE - For use with high throughput sequencing technologies, Whole Genome Amplification kit designed to facilitate Next Gen Sequencing.
Synonym: SeqPlex Enhanced DNA Amplification Kit; WGA kit; whole genome amplification kit
Product Type: Chemical
The SeqPlex Enhanced DNA Amplification Kit (SEQXE) was used to amplify 1 μL of ChIP-selected DNA for comparison with unamplified ChIP DNA in Illumina sequencing. Three separate ChIP selections were performed with Sigma’s Imprint® Ultra ChIP kit (CHP2) each using 5 million BJ fibroblast cells and 0.5–2 μg H3K9me3-specific antibody (SAB4800004). Each ChIP selection was eluted in 20 μL of CHP2 DNA Elution Buffer (B5688) after the final purification. The three ChIP preparations were pooled and 1 μL (±0.2 ng) was amplified according to the procedure described in the Technical Bulletin for the SeqPlex Enhanced DNA Amplification Kit. After primer removal, the SeqPlex DNA and 20 μL from the same starting pool of ChIP DNA were both treated to standard Illumina library preparation and sequenced on an Illumina MiSeq. Excellent correlation is seen between SeqPlex amplified and Unamplified reads in this view of greater than 1 million base pairs on Chromosome 21.
Closer inspection also reveals similar sequence alignments for SeqPlex (top) and Unamplified (bottom). Both sets of Chromosome 21 reads were visualized using the Integrated Genome Browser 6.7.3.
SeqPlex is able to amplify nearly all (97%) of the peaks detected in the Unamplified sample. Up to 60% of the potential regions of interest would have been missed without amplification.
Very similar results are achieved each time a sample is amplified. Above is a Peak Correlation plot showing excellent reproducibility for two different amplifications from the same ChIP sample.
| Quality Level | 200  |
| shipped in | wet ice |
| storage temp. | −20°C |
| technique(s) | whole genome amplification: suitable |
| Application: | SeqPlex™ DNA Amplification Kit has been used for whole genome amplification. |
| Features and Benefits: | • Random priming technology amplifies fragmented DNA such as ChIP or FFPE • Facilitates sequencing from as little as 100 pg of ChIP DNA • Enhanced primers for complete genome coverage, minimal sequence bias, primer removal, and amplicon size ideal for next gen sequencing • Compatible with Illumina®, SOLiD™, or 454 library prep for next generation sequencing |
| General description: | SeqPlex™ Enhanced DNA Amplification Kit for whole genome amplification (WGA) is designed to facilitate next-generation sequencing (NGS) from extremely small quantities or from degraded/highly fragmented DNA. The yields from chromatin immunoprecipitation (ChIP) or formalin-fixed paraffin-embedded tissue samples (FFPE) are often less than required for successful next generation sequencing library preparation. The SeqPlex kit allows the user to pre-amplify these and other small quantity/highly fragmented DNA samples for input into a NGS workflow. This kit is an extension of the WGA product line and has been developed to integrate into the Illumina®, SOLiD™, or 454 sequencing workflows. |
| Legal Information: | iCAT is a registered trademark of University of Washington |
| Legal Information: | Illumina is a registered trademark of Illumina, Inc. |
| Legal Information: | SeqPlex is a trademark of Sigma-Aldrich Co. LLC |
| Legal Information: | SOLiD is a trademark of Applera Corporation or its subsidiaries in the US and/or certain other countries |
| Other Notes: | SEQXE-500RXN is manufactured on-demand. Contact technical services at techserv@sial.com for more information. |
| RIDADR | NONH for all modes of transport |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| Storage Temp. | −20°C |
| UNSPSC | 12352200 |
| Components | Water, Nuclease-Free Water, for Molecular Biology; Library Preparation Buffer |