The kit contains two types of cDNA 2nd Strand synthesis buffer, which can be chosen for library
construction for non-stranded or stranded RNA-Seq transcriptome analysis.
This kit combines 2nd Strand cDNA synthesis, end-repair and dA-Tailing into one step, with no need of purification, which greatly simplifies the process
of library construction and shortens the operation time. The optimized reaction system improves the library construction efficiency, is compatible with
lower-input RNA, and has uniform coverage for different amounts of input-RNA.
Libraries of specific sizes, which can be customized, can be obtained after size selection with magnetic beads. All the enzymes and buffer provided in
the kit have undergone rigorous quality control and functional testing to ensure the optimal stability and repeatability.
VAHTS Universal V6 RNA-seq Library Prep Kit for Illumina is suitable for RNA library construction of RNA that have been enriched by Poly(A) (for RNA
with good integrity from eukaryotes such as animals, plants and fungi) or rRNA depletion. As the content of mRNA in total RNA of different samples
varies greatly, enough total RNA need to be inputted to make sure the sufficient mRNA for library construction. The amount of input-RNA is related to
the mRNA enrichment module:
VAHTS mRNA Capture Beads (Vazyme #N401): 0.01 - 4 μg;
Ribo-off rRNA Depletion Kit (H/R/M) (Vazyme #N406): 0.01 - 1 μg;
Ribo-off rRNA Depletion Kit (Bacteria) (Vazyme #N407): 0.01 - 5 μg;
Ribo-off Globin & rRNA Depletion Kit (H/M/R) (Vazyme #N408): 0.01 - 1μg;
Ribo-off rRNA Depletion Kit (Plant) (Vazyme #N409): 1 - 5 μg;
Purified mRNA or Ribosomal-depleted RNA: 0.5 - 100 ng;
It is recommended to use Agilent 2100 Bioanalyzer to analyze the integrity of total RNA. mRNA enriched with VAHTS mRNA Capture Beads
(Vazyme #N401) must be high quality RNA (RIN ≥ 7). Degraded total RNA used for library construction will lead to 3’ bias in RNA-seq. For RNA
samples with RIN value < 7, rRNA removal can be performed using the Ribo-off method (Vazyme #N406/407/408/409).
Main fields of RNA-related analysis:
◇ gene expression analysis
◇ single nucleotide variation calling