Merck SEQPLEX-I WTA Kit

SEQPLEX-I WTA Kit

Whole Transcriptome Amplification (RNA Amplification)

기술 적용

• Whole genome amplification: suitable
• Whole transcriptome amplification: suitable

호환성

• Illumina (Next Generation Sequencing)

배송 상태

• Wet ice

저장 온도

• −20°C

제품 설명

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: Pre-amplification / Library Synthesis

Using Library Preparation Reagents, template RNA is reverse transcribed with primers composed of semi-degenerate 3′ and universal 5′ ends.
During polymerization, displaced and RNaseH-generated single strands serve as new templates for additional primer annealing and extension, producing random overlapping cDNAs flanked by universal primer (5′) and primer complement (3′) sequences.

Step 2: Amplified Library Synthesis

Using Amplification 1 Reagents, products from pre-amplification/library synthesis are amplified by single primer PCR via the universal end sequence.
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 depend on the length of the starting RNA.

Step 3: Sequencing Library Synthesis

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.

제품 사양