Small RNA Library Prep Kit for Illumina

Figure 2. An example of a small RNA library on resolved on a 6% Novex® TBE PAGE gel. The library containing microRNAs resolved at around 140 bp and migrated just below the 150 bp marker of the provided NGS MW Ladder. The NGS Control Ladder contains two bands for alignment during band excision. The top band co-migrate with the expected size of library construct containing mainly microRNAs (~140 bp). The bottom band co-migrate with the expected size of an adaptor-adaptor-ligation product (~ 120 bp) that should be excluded.

Figure 3. Superb reproducibility of constructing small RNA (microRNA) sequencing libraries from liquid biopsies samples with low RNA abundance. Total RNA was prepared from 200 µL of EDTA-plasma using Norgen's Plasma/Serum RNA Purification Mini Kit (Cat. 55000). Small RNA libraries (four replicates) were prepared from purified RNA using Norgen’s Small RNA Library Prep Kit for Illumina (Cat. 63600) without the use of gel purification (prep time of ~ 5 hours). The resulting libraries were sequenced on Illumina's MiSeq system using v3 chemistry. Reads were mapped to microRNAs and normalized to reads per million. Pairwise-comparisons of microRNA expression among all replicates were performed. High correlations (or reproducibility) were observed among all replicates with an average of over 99%.

Figure 4. Combining true total RNA purification technology with robust production of small RNA (microRNA) sequencing libraries from RNA purified from whole blood and preserved blood. Total RNA was prepared from either whole blood or collected in Paxgene™ or Tempus™ blood tubes using Norgen’s Total RNA Purification Plus Kit (Cat. 48300), Preserved Blood RNA Purification Kit II (for use with Paxgene™ Blood RNA Tubes) (Cat. 43500) or Preserved Blood RNA Purification Kit I (for use with Tempus™ Blood RNA Tubes) (Cat. 43400), respectively, in duplicates. Small RNA libraries were prepared from purified RNA using Norgen's Small RNA Library Prep Kit for Illumina (Cat. 63600). The resulting libraries were sequenced on Illumina's MiSeq system using v3 chemistry. Reads were mapped to microRNAs and normalized to reads per million. Pairwise-comparisons of microRNA expression among all replicates were performed. High correlations (or reproducibility) were observed among all replicates of each blood tube with an average of over 99% (Panel A). Moreover, all three types of blood samples yielded similar numbers of microRNA detected (Panel B) with highly correlated expression profiles (Panel C)