VAHTS Universal V8 RNA-seq Library Prep Kit for Illumina is used for preparing transcriptome libraries for Illumina platform. The kit is universal and appropriate for RNA library construction of RNA that have been obtained by Poly(A)-based mRNA enrichment or rRNA depletion.
The kit comprises two types of cDNA 2nd Strand synthesis buffer. The buffers can be selected for library construction for stranded or non-stranded RNA-Seq transcriptome analysis. The kit integrates 2nd Strand cDNA synthesis, dA-Tailing, and end-repair into one step, eliminating the need for purification, thus simplifying the library construction process and reducing the operation time.
The optimized reaction system enhances the library construction efficiency, has uniform coverage for varying amounts of input-RNA, and is compatible with lower-input RNA. It is possible to obtain customized libraries of specific sizes after size selection with magnetic beads. Strict quality control and functional testing is done for the kit’s enzymes and buffer to achieve optimal stability and repeatability.
Image Credit: Nanjing Vazyme Biotech Co., Ltd.
- High success rate
- Wide-ranging species compatibility
- Compatibility: Better success rate of library construction for low initial volume and degraded samples
- Quick: Simple operation process, which shortens the time of building the transcription library to 3 hours
- High quality: Better sequencing data quality, resulting in a more even coverage of the transcript region
VAHTS Universal V8 RNA-seq Library Prep Kit for Illumina can be used for RNA library construction of total RNA that is enriched by Poly(A) (for RNA with good integrity from eukaryotes such as plants, animals and fungi) or rRNA depletion. Due to the large variation in the content of mRNA in total RNA of different samples, sufficient total RNA needs to be inputted to ensure sufficient mRNA for library construction.
The amount of input-RNA depends on the mRNA enrichment module:
Ribo-off rRNA Depletion Kit (Human/Mouse/Rat) (Vazyme #N406): 0.01–1 g; VAHTS mRNA Capture Beads (Vazyme #N401): 0.01–4 g; Ribo-off rRNA Depletion Kit (Bacteria) (Vazyme #N407): 0.01– 5 g; Ribo-off rRNA Depletion Kit (Plant) (Vazyme #N409): 1–5 g; Ribo-off Globin & rRNA Depletion Kit (Human/Mouse/Rat) (Vazyme #N408): 0.01–1 g; Purified mRNA or Ribosomal-depleted RNA: 0.5–100 ng.
It is preferable to use Agilent 2100 Bioanalyzer to study the integrity of total RNA. mRNA supplemented with VAHTS mRNA Capture Beads (Vazyme #N401) should be of high-quality RNA (RIN 7) because degraded total RNA used for library construction will result in a 3' bias in RNA-seq. For RNA samples with RIN value <7, the Ribo-off method (Vazyme #N406/407/408/409) is suitable for rRNA removal.
Main fields of RNA-related analysis:
- Gene expression analysis
- Alternative splicing detection
- Single nucleotide variation calling
- Target transcriptome analysis
- Gene fusion detection
- RNA analysis: Equalbit RNA BR Assay Kit (Vazyme #EQ212); Equalbit RNA HS Assay Kit (Vazyme #EQ211); Agilent RNA 6000 Pico Kit (Agilent #5067-1513).
- mRNA enrichment & rRNA depletion: Ribo-off rRNA Depletion Kit (Human/Mouse/Rat) (Vazyme #N406); VAHTS mRNA Capture Beads (Vazyme #N401); Ribo-off rRNA Depletion Kit (Bacteria) (Vazyme #N407); Ribo-off rRNA Depletion Kit (Plant) (Vazyme #N409); Ribo-off Globin & rRNA Depletion Kit (Human/Mouse/Rat) (Vazyme #N408).
- RNA and DNA clean beads: VAHTS RNA Clean Beads (Vazyme #N412) or Agencourt RNA Clean XP Beads (Beckman #A63987); VAHTS DNA Clean Beads (Vazyme #N411) or Agencourt AMPure XP Reagent (Beckman #A63880/A63881/A63882).
- 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: Agilent DNA 1000 Kit (Agilent #5067-1504) or Agilent High Sensitivity DNA Kit (Agilent #5067 - 4626); Equalbit 1 × dsDNA HS Assay Kit (Vazyme #EQ121).
- Other materials: Freshly prepared Ethanol (80%), Nuclease-free ddH2O, Low-adsorption Nuclease-free PCR Tubes and Pipette Tips, PCR Instrument, Magnetic Stand, Qubit, Agilent 2100 Bioanalyzer.