Apr 3, 2016

Evaluation of TagSeq, a reliable low-cost alternative for RNAseq

Molecular Ecology Resources
Brian K LohmanDaniel I Bolnick


RNAseq is a relatively new tool for ecological genetics that offers researchers insight into changes in gene expression in response to a myriad of natural or experimental conditions. However, standard RNAseq methods (e.g., Illumina TruSeq(®) or NEBNext(®) ) can be cost prohibitive, especially when study designs require large sample sizes. Consequently, RNAseq is often underused as a method, or is applied to small sample sizes that confer poor statistical power. Low cost RNAseq methods could therefore enable far greater and more powerful applications of transcriptomics in ecological genetics and beyond. Standard mRNAseq is costly partly because one sequences portions of the full length of all transcripts. Such whole-mRNA data are redundant for estimates of relative gene expression. TagSeq is an alternative method that focuses sequencing effort on mRNAs' 3' end, reducing the necessary sequencing depth per sample, and thus cost. We present a revised TagSeq library construction procedure, and compare its performance against NEBNext(®) , the 'gold-standard' whole mRNAseq method. We built both TagSeq and NEBNext(®) libraries from the same biological samples, each spiked with control RNAs. We found that TagSeq measured the control RNA...Continue Reading

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  • Citations32


Mentioned in this Paper

Sequence Determinations, RNA
Research Personnel
Nucleic Acid Sequencing
JM 2820
Gene Expression
Gold Measurement
Molecular Genetic Technique
Surgical Construction

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