Merck SEQPLEX-I WTA Kit

SEQPLEX-I WTA Kit

Whole Transcriptome Amplification, RNA Amplification

technique(s)

whole genome amplification: suitable
whole transcriptome amplification: suitable

호환성

Illumina (Next Generationa Sequencing)

배송 상태

wet ice

저장 온도

−20°C

The SeqPlex™-I WTA kit allows amplification of small quantities of reverse transcribed RNA or degraded RNA for direct input onto Illumina® next-generation sequencing (NGS) flow cells. The SeqPlex-i process is comprised of three steps: Pre-amplification/Library Synthesis, Amplification 1 and Amplification 2

Step 1: In the Pre-amplification/Library Synthesis step using the (Library Preparation Reagents), the template RNA is reverse transcribed using primers composed of a semi-degenerate 3′- and universal 5′-ends. As polymerization proceeds, displaced and RNaseH generated single strands serve as new templates for additional primer annealing and extension producing random, overlapping cDNAs flanked by a universal primer (5′) and primer complement (3′) sequence.

Step 2: In the Amplified Library Synthesis step (using the Amplification 1 Reagents), products from pre-amplification/library synthesis are amplified by single primer PCR via the universal end sequence. These amplification 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 (using Amplification 2 Reagents), single primer amplicons from amplification 1 are converted to dual Illumina® primer PCR products ready for purification, quantification, and Illumina® NGS.