Optimization of illumina Truseq Targeted RNA expression (TREx) library quality
RNA-seq has become an essential tool in molecular research. Nevertheless, application of RNA-seq was limited by cost and technical difficulties. Illumina has introduced the cost effective and ease to handle Truseq Targeted RNA Sequencing. In this study, we present the requirements and the optimiza...
Main Authors: | , , , , , |
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Format: | Article |
Language: | English |
Published: |
Penerbit Universiti Kebangsaan Malaysia
2018
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Online Access: | http://journalarticle.ukm.my/12009/ http://journalarticle.ukm.my/12009/ http://journalarticle.ukm.my/12009/1/UKM%20SAINSMalaysiana%2047%2802%29Feb%202018%20%2012.pdf |
Summary: | RNA-seq has become an essential tool in molecular research. Nevertheless, application of RNA-seq was limited by cost and
technical difficulties. Illumina has introduced the cost effective and ease to handle Truseq Targeted RNA Sequencing. In
this study, we present the requirements and the optimization procedure for this Truseq Targeted RNA sequencing on cell
line. Total RNA was recommended as starting materials but it required optimization including additional purification
step and adjusting the AMPure beads ratio to eliminate unwanted contaminants. This can be resolved by using PolyAenriched
mRNA as starting material. TREx is a useful assay to evaluate gene expression. Quality library of TREx can be
prepared by adding multiple washing steps or changing input sample to mRNA. |
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