Bioo Scientific announced the release of the NEXTflex directional RNA-Seq kits and barcodes primer sets which provide an easy, flexible and non-biased solution for generating directional single, paired-end and multiplexed libraries from mRNA or rRNA depleted total RNA.
Using the NEXTflex Directional RNA-Seq Kit, RNA can be sequenced in the 5’-3’ and the 3’-5’ direction thus allowing for thorough and even coverage of long RNAs. Using the NEXTflex Directional RNA-Seq Kits, mRNA or rRNA depleted RNA is fragmented using a cationic buffer. Fragmented RNA is then dephosphorylated, followed by a kinase treatment which adds a 5’phosphate. Fragmented RNA now containing 5’phosphate and 3’hydroxyl groups are then ligated to randomized 3’ and 5’ adapters followed by first strand synthesis and amplification.
Bidirectional sequencing can be performed on the same sample by using barcodes to differentiate the different reads.
While ligation based adapter addition is a hallmark of most NGS library preparations, it is well known that ligases can introduce sequence dependent bias, an un-desirable side effect. Using Bioo Scientific’s patent pending randomization technology, mixed bases are incorporated at the ends of the adapter eliminating ligase preference and its resulting bias. This technology is available in both NEXTflex Randomized Adapter and NEXTflex Randomized Reverse Adapters Sets.
The NEXTflex Randomized Adapters in the NEXTflex Directional RNA-Seq Kit are incorporated to eliminate ligation associated biases thus increasing the accuracy of transcriptome profiles.