The VAHTSTM Total RNA-seq (H / M / R) Library Prep Kit for Illumina® is a special strand-specific transcriptome library construction kit for the Illumina high-throughput sequencing platform. This kit is suitable for the amount of starting templates. The total RNA of humans, mice and rats is 0.1 to 1 µg. The difference from the conventional transcriptome library is that this kit can combine rRNA (including cytoplasmic rRNA 28S, 18S, 5S and mitochondrial rRNA 16S, 12S, 5.8S). ) It is removed from total RNA, leaving mRNA and other non-coding RNAs.
It is used for the analysis of non-coding RNA, such as LncRNA; Degraded RNA samples can also be used for library construction with this kit. When the second-strand cDNA is synthesized in this kit, dUTP is added to label it and the second tagged strand is digested with the UDG enzyme prior to PCR, so the sequencing information comes from the first-strand cDNA, resulting which ensures the directionality of the strand. RNA Analysis Analyze the sequencing data to determine whether the transcript is from the sense or antisense DNA strand.
The kit includes three independent packages of NR4, NR5 and NR6, which contain the library construction process.
All necessary enzymes, buffers, and components have undergone strict quality control and functional verification, ensuring the stability and repeatability of the library construction to the greatest extent possible. The DNA and RNA purification magnetic beads and adapters must be prepared separately.
Adapters can be selected from VAHTSTM RNA Adapters (Vazyme # N803, Vazyme # N804 or Vazyme # N809, Vazyme # N810, Vazyme # N811, Vazyme # N812). For details on the adapter selection, see Common problems and solutions 6 (Page 17); Magnetic DNA Purification Beads can choose from VAHTSTM DNA Clean Beads (Vazyme # N411) or Agencourt® AMPure® XP Reagent (Beckman # A63880, # A63881, # A63882); RNA Purification Magnetic Beads can be chosen from VAHTSTM RNA Clean Beads (Vazyme # N412) or Agencourt® RNAClean® XP Beads (Beckman # A63987).
Fast: minimalist operating process reduces database creation time by 2 h
Compatibility: compatible with a variety of RNA enrichment modules, with any combination
General: flexible and free choice of chain-specific or non-chain-specific libraries
Storage: All components are stored at -20 ℃