2. Transcriptome Analysis:
Library Construction
Hsin-Yi Chang and Hsueh-Fen Juan∗
Institute of Molecular and Cellular Biology,
National Taiwan University, Taipei, Taiwan
∗[email protected]- Introduction
Transcriptome analysis reveals pivotal gene expression regulations in given
biological statuses. Conventional microarray analyses provide opportunities
to study relevant levels of transcripts with known sequences.^1 Therefore, gene
expressions, alternative splicing of genes, and amount of any user-defined
sequences can be measured in high-throughput manner. However, several
limitations arise while employing microarray analysis: Loss of linearity of
signal, limited detection of weak fluorescence intensity, and inability to
discover novel transcripts.2–
Next-generation sequencing (NGS) provides digital counts in transcripts,
sequence information in single nucleotide resolution, and high sensitive
detection in single read. These advantages give rise to a step forward
towards revealing new phenotypes of transcriptome. Transcriptome analysis
has been successfully applied rapidly in various biological issues, such as gene
expression, differential splicing, and allele-specific expression of transcripts.
Furthermore, the application of NGS in detecting specific diseases, specific
genes or mutations (targeted sequencing) for pathological determination
makes the implementation in personalized medicine progressive.5,6Moreover,
the advantage of NGS in sequencing non-model organisms is to provide the
analytical power to reveal gene structures rapidly and molecular ecology
discipline.7,
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