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 optimizati...

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Main Authors: Abu, Nadiah, Yeap, Swee Keong, Nordin, Noraini, Tan, Sheau Wei, Ho, Wan Yong, Mohammed Alitheen, Noorjahan Banu
Format: Article
Language:English
Published: Penerbit Universiti Kebangsaan Malaysia 2018
Online Access:http://psasir.upm.edu.my/id/eprint/60474/
http://psasir.upm.edu.my/id/eprint/60474/1/Optimization%20of%20illumina%20TruSeq%20Targeted%20RNA%20expression%20%28TREx%29%20library%20quality.pdf
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author Abu, Nadiah
Yeap, Swee Keong
Nordin, Noraini
Tan, Sheau Wei
Ho, Wan Yong
Mohammed Alitheen, Noorjahan Banu
author_facet Abu, Nadiah
Yeap, Swee Keong
Nordin, Noraini
Tan, Sheau Wei
Ho, Wan Yong
Mohammed Alitheen, Noorjahan Banu
author_sort Abu, Nadiah
building UPM Institutional Repository
collection Online Access
description 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 PolyA-enriched 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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institution Universiti Putra Malaysia
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language English
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publisher Penerbit Universiti Kebangsaan Malaysia
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spelling upm-604742018-05-21T03:51:08Z http://psasir.upm.edu.my/id/eprint/60474/ Optimization of illumina TruSeq Targeted RNA expression (TREx) library quality Abu, Nadiah Yeap, Swee Keong Nordin, Noraini Tan, Sheau Wei Ho, Wan Yong Mohammed Alitheen, Noorjahan Banu 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 PolyA-enriched 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. Penerbit Universiti Kebangsaan Malaysia 2018 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/60474/1/Optimization%20of%20illumina%20TruSeq%20Targeted%20RNA%20expression%20%28TREx%29%20library%20quality.pdf Abu, Nadiah and Yeap, Swee Keong and Nordin, Noraini and Tan, Sheau Wei and Ho, Wan Yong and Mohammed Alitheen, Noorjahan Banu (2018) Optimization of illumina TruSeq Targeted RNA expression (TREx) library quality. Sains Malaysiana, 47 (2). pp. 303-308. ISSN 0126-6039 http://www.ukm.my/jsm/english_journals/vol47num2_2018/contentsVol47num2_2018.html 10.17576/jsm-2018-4702-12
spellingShingle Abu, Nadiah
Yeap, Swee Keong
Nordin, Noraini
Tan, Sheau Wei
Ho, Wan Yong
Mohammed Alitheen, Noorjahan Banu
Optimization of illumina TruSeq Targeted RNA expression (TREx) library quality
title Optimization of illumina TruSeq Targeted RNA expression (TREx) library quality
title_full Optimization of illumina TruSeq Targeted RNA expression (TREx) library quality
title_fullStr Optimization of illumina TruSeq Targeted RNA expression (TREx) library quality
title_full_unstemmed Optimization of illumina TruSeq Targeted RNA expression (TREx) library quality
title_short Optimization of illumina TruSeq Targeted RNA expression (TREx) library quality
title_sort optimization of illumina truseq targeted rna expression (trex) library quality
url http://psasir.upm.edu.my/id/eprint/60474/
http://psasir.upm.edu.my/id/eprint/60474/
http://psasir.upm.edu.my/id/eprint/60474/
http://psasir.upm.edu.my/id/eprint/60474/1/Optimization%20of%20illumina%20TruSeq%20Targeted%20RNA%20expression%20%28TREx%29%20library%20quality.pdf