SEQPLEX-I WGA Kit
SIGMA/SEQXI - Whole Genome Amplification, DNA Amplification
Synonym: SeqX-i DNA Amplification kit; WGA Kit; Whole Genome Amplification kit
Product Type: Chemical
| compatibility | Illumina Next Generation Sequencing |
| shipped in | wet ice |
| storage temp. | −20°C |
| technique(s) | DNA amplification: suitable |
| PCR: suitable | |
| whole genome amplification: suitable |
| Application: | SeqPlex™-I WGA Kit has been used for whole genome amplification. |
| Disclaimer: | The SeqPlex-I DNA Amplification Kit for whole genome amplification (WGA) is for R&D use only. Not for drug, household, or other uses. |
| Features and Benefits: | • Amplifies fragmented/extremely small quantities of DNA: Yields from ChIP or FFPE, which are 200-500 bp are easily amplified by Random priming technology. • Facilitates sequencing from as little as 100 pg of DNA • Enhanced primers for complete genome coverage, minimal sequence bias, and amplicon size ideal for next generation sequencing (NGS) • Cost-effective: No longer requires an additional NGS library prep step • Compatible with Illumina® next-generation sequencing |
| General description: | The SeqPlex™-I WTA kit allows amplification of smallquantities of reverse transcribed RNA or degraded RNA for direct input ontoIllumina® next-generation sequencing (NGS) flow cells. TheSeqPlex-i process is comprised of three steps: Pre-amplification/Library Step 1: In the Pre-amplification/Library Synthesis stepusing the (Library Preparation Reagents), the template RNA is reversetranscribed using primers composed of a semi-degenerate 3′- and universal5′-ends. As polymerization proceeds, displaced and RNaseH generated singlestrands serve as new templates for additional primer annealing and extensionproducing random, overlapping cDNAs flanked by a universal primer (5′) andprimer complement (3′) sequence. Step 2: In the Amplified Library Synthesis step (using theAmplification 1 Reagents), products from pre-amplification/library synthesisare amplified by single primer PCR via the universal end sequence. Theseamplification products typically range from 200 to 500+ base pairs. Note:Amplicons from degraded RNA, such as Formalin Fixed Paraffin Embedded (FFPE),are typically shorter and dependent upon the length of the starting RNA. Step 3: In the Sequencing Library Synthesis step (usingAmplification 2 Reagents), single primer amplicons from amplification 1 areconverted to dual Illumina® primer PCR products ready forpurification, quantification, and Illumina® NGS. |
| General description: | To order SeqPlexI adapters in tubes, please download NGSO Adapters SeqPlexI tubes (XLSX File)  . To order arrayed adapters in plates, download NGSO Plate SeqPlexI arrayed plates (XLSX File) . Details on our custom adapters product, Next-Gen Sequencing Oligos (NGSO), can be found at SigmaAldrich.com/nextgeno |
| Legal Information: | Illumina is a registered trademark of Illumina, Inc. |
| Legal Information: | SeqPlex is a trademark of Sigma-Aldrich Co. LLC |
| Other Notes: | • A 20 μL Amplification 2 reaction will produce >100 ng of amplified double-stranded cDNA when starting with 100 pg to 5 ng of high-quality DNA. Higher input quantities and higher quality DNA template generally result in increased yields. For damaged DNA, such as from FFPE, 1–50 ng input DNA is recommended. • The dual index adapter primers (AP100) provided in this kit will only work for one sample. If pooling of samples for sequencing is required, the user must provide additional index primer sets. See example index primer sequences on page 2 of the technical bulletin. |
| Storage Temp. | −20°C |
| UNSPSC | 41121800 |