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Location: Home > Custom Services > Sequencing and Chip Technology > Massive Parallel Signature Sequencing

Massive Parallel Signature Sequencing

Date: 2018-01-25 Author: Leading Biology Click: 880

Introduction

Massive Parallel Signature Sequencing (MPSS) is a method for determining expression levels of mRNA by counting the number of individual mRNA molecules produced by each gene. During MPSS, A sample of mRNA is first converted to complementary DNA (cDNA) using reverse transcriptase, which makes subsequent manipulations easier. These cDNAs are fused to a small oligonucleotide "tag" which allows the cDNA to be PCR amplified and then coupled to microbeads. After several rounds of sequence determination, using the hybridization of fluorescent-labeled probes, a sequence signature of ~16-20 bp is determined from each bead. Fluorescent imaging captures the signal from all the beads, so DNA sequences are determined from all the beads in parallel. 


Each signature sequence in a MPSS dataset is analyzed, compared with all other signatures and all identical signatures are counted. The level of expression of any single gene is calculated by dividing the number of signatures from that gene by the total number of signatures for all mRNAs present in the dataset. MPSS datasets are additive in nature, which means that datasets from multiple analyses with the same starting mRNA sample can be combined.


MPSS has routine sensitivity at a level of a few molecules of mRNA per cell, and the datasets are in a digital format that simplifies the management and analysis of the data.


Massive Parallel Signature SequencingMassive Parallel Signature Sequencing

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