SEQPLEX-I WTA Kit

REF : SEQRI-96RXN
Marca : Sigma-Aldrich
Descrição :SEQPLEX-I WTA Kit Whole Transcriptome Amplification, RNA Amplification
Clique para mais informações
Categoria :
Descrição detalhada : The SeqPlex(TM)-I WTA kit allows amplification of small quantities of reverse transcribed RNA or degraded RNA for direct input onto Illumina(R) next-generation sequencing (NGS) flow cells. The SeqPlex-i process is comprised of three steps: Pre-amplification/Library Synthesis, Amplification 1 and Amplification 2Step 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(R) primer PCR products ready for purification, quantification, and Illumina(R) NGS.
Armazenamento : -20C
Embalagem : 1X96RXN