The Vazyme VAHTS Total RNA-seq (H/M/R) Library Prep Kit for Illumina is specially designed for the preparation of stranded transcriptome libraries for next generation sequencing (NGS) platforms of Illumina. The initial template is prepared with 0.1 µg-1 µg of total RNA of
human, mouse, or rat. Compared with traditional transcriptome library preparation kits, this kit enables the depletion of ribosomal RNA
(rRNA), including both cytoplasmic (28S, 18S, 5.8S, and5S) and mitochondrial (16S and 12S) rRNA, leaving mRNA and other non-coding
RNA behind. This kit is suitable for subsequent non-coding RNA analysis such as lncRNA. Degraded RNA (i.e. FFPE RNA) could also be
used to prepare the library with this kit. In addition, this kit enables the insertion of dUTP during the 2nd strand synthesis of cDNA. The
double strand cDNA are digested by uracil-DNA glycosylase (UDG) to remove the second strand before sequencing. As a result, only
information from the 1st strand cDNA is preserved. In addition to standard transcriptome information, strand-specific (i.e. from sense or
anti-sense DNA) information can also be obtained from NGS data analysis.
heat labile udg vazyme
Applications
Requirements for Starting Materials: 0.1 µg-1 µg total RNA of human, mouse. Degraded RNA (i.e. FFPE RNA) could also be used to
prepare the library with this kit.
Information of Transcripts: : This kit is applicable to stranded mRNA or non-coding RNA (with the exception of rRNA) related analysis
with RNA-seq, including gene expression analysis, single nucleotide variation calling, alternative splicing / fusion detection, target
transcriptome analysis, and target genes prediction and functional analysis.
If conventional mRNA of animal, plant, or fungal is under consideration, please use the VAHTS mRNA-seq V2 Library Prep Kit for Illumina
(Vazyme, #NR601) for library construction. If stranded mRNA of animal, plant, or fungal is under consideration, please use the VAHTS
Stranded RNA-seq Library Prep Kit for Illumina (Vazyme, #NR602) for library construction.
Additional Materials Required
DNA Clean Beads: VAHTS DNA Clean Beads (Vazyme,#N411) or Agencourt MPure XP Beads (Beckman Coulter, #A63880, #A63881,
#A63882).
RNA Clean Beads: VAHTS RNA Clean Beads (Vazyme, #N412) or Agencourt RNAClean XP Beads(Beckman Coulter, #A63987).
RNA Analysis：Agilent RNA 6000 Pico Kit (Agilent, #5067-1513).
Library Analysis: Agilent DNA 1000 Kit (Agilent, #5067-1504).
Adapters: VAHTS RNA Adapters Set 1 – Set 2 for Illumina (Vazyme, #N803, #N804), or VAHTS RNA Adapters Set 3 – Set 6 for Illumina
(Vazyme, #N809, #N810, #N811, #N812).
Other Materials: Fresh Ethanol (80%), Nuclease-free Water, Nuclease-free PCR tubes, Low absorption EP tubes, Agilent 2100 Bioanalyzer, Thermocyler (PCR instrument) ,Magnetic stand.
VAHTS Universal V6 RNA-seq Library Prep Kit for Illumina is specially designed for the preparation of transcriptome libraries for next generation
sequencing (NGS) platforms of Illumina. The kit is universal and suitable for RNA library construction of RNA that have been obtained by
Poly(A)-based mRNA enrichment or rRNA depletion . The kit contains two types of cDNA 2nd Strand synthesis buffer, which can be chosen for library
construction for non-stranded transcriptome or stranded 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 selections with magnetic beads.
01/ Introduction
All Components should be stored at -30℃ ~ -15℃ , transported at -20℃ ~ 0℃.
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): 0.05μg – 4 μg;
Ribo-off rRNA Depletion Kit (H/R/M) (Vazyme #N406): 0.05μg – 1 μg;
Ribo-off rRNA Depletion Kit (Bacteria) (Vazyme #N407): 1 μg – 5 μg;
Ribo-off rRNA Depletion Kit (Plant) (Vazyme #N409): 1 μg – 5 μg;
It is recommended to use an Agilent 2100 Bioanalyzer to analyze the integrity of total RNA. mRNA enrichment using VAHTS® mRNA Capture Beads
(Vazyme #N401) needs high quality RNA samples (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 #N406/407/409).
Main fields of RNA-related analysis:
◇gene expression analysis
◇single nucleotide variation calling
◇alternative splicing detection
◇gene fusion detection
◇target transcriptome analysis
04/ Applications
02/ Components
03/ Storage
