VAHTS Universal V8 RNA-seq Library Prep Kit for Illumina

VAHTS Universal V8 RNA-seq Library Prep Kit for Illumina is specially designed for the preparation of transcriptome libraries for Illumina platform. 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 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.

Product advantages

• Fast: the operation process is very simple, which reduces the time of building the transcription library to 3 h
• Compatibility: higher success rate of library construction for low initial volume and degraded samples
• High quality: Excellent sequencing data quality, resulting in more uniform coverage of the transcript region

Applications

VAHTS Universal V8 RNA-seq Library Prep Kit for Illumina is suitable for RNA library construction of total RNA that have been enriched by Poly(A) (for RNA with good integrity from eukaryotes such as animals, plants nd 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 (Human/Mouse/Rat) (Vazyme #N406): 0.01 - 1 μg;
• Ribo-off rRNA Depletion Kit (Bacteria) (Vazyme #N407): 0.01 - 5 μg;
• Ribo-off Globin & rRNA Depletion Kit (Human/Mouse/Rat) (Vazyme #N408): 0.01 - 1 μg;
• 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
• alternative splicing detection
• gene fusion detection
• target transcriptome analysis

Self-prepared materials

RNA Analysis:

• Equalbit RNA HS Assay Kit (Vazyme #EQ211);
• Equalbit RNA BR Assay Kit (Vazyme #EQ212);
• Agilent RNA 6000 Pico Kit (Agilent #5067-1513).
• mRNA Enrichment & rRNA Depletion:

VAHTS mRNA Capture Beads (Vazyme #N401);

• Ribo-off rRNA Depletion Kit (Human/Mouse/Rat) (Vazyme #N406);
• Ribo-off rRNA Depletion Kit (Bacteria) (Vazyme #N407);
• Ribo-off Globin & rRNA Depletion Kit (Human/Mouse/Rat) (Vazyme #N408);
• Ribo-off rRNA Depletion Kit (Plant) (Vazyme #N409).

RNA and DNA Clean Beads:

• VAHTS DNA Clean Beads (Vazyme #N411) or Agencourt AMPure XP Reagent
• (Beckman #A63880/A63881/A63882);
• VAHTS RNA Clean Beads (Vazyme #N412) or Agencourt RNA Clean XP Beads
• (Beckman #A63987).

Adapters:

• VAHTS RNA Adapters Set 1 - Set 2 for Illumina (Vazyme #N803/N804), VAHTS RNA
• Adapters Set 3 - Set 6 for Illumina (Vazyme #N809/N810/N811/N812) or VAHTS RNA
• Multiplex Oligos Set 1 - Set 2 for Illumina (Vazyme #N323/N324).

Library Quality Control:

• Equalbit 1 × dsDNA HS Assay Kit (Vazyme #EQ121);
• Agilent DNA 1000 Kit (Agilent #5067-1504) or Agilent High Sensitivity DNA Kit (Agilent #5067 - 4626).

Other Materials:

• Freshly prepared Ethanol (80%), Nuclease-free ddH2O, Low-adsorption Nuclease-free PCR Tubes andPipette Tips, PCR Instrument, Magnetic Stand, Qubit, Agilent 2100 Bioanalyzer.