NEXTflex Small RNA Sequencing Kit for Library Prep Launched

[Bioo Scientific]

Bioo Scientific launched the patent pending NEXTflex™ Small RNA Sequencing Kit which provides an easy, flexible, cost-effective solution for generating single and multiplexed libraries from small RNA using Illumina GAIIx and HiSeq 2000 sequencing platforms. The NEXTflex Small RNA Sequencing Kit allows users to generate libraries directly from as little as 1 μg of total RNA. This kit contains a specialized 3’ adenylated adapter that specifically ligates onto microRNAs and other small RNAs containing a 3’ hydroxyl group. After adapter ligation and reverse transcription, libraries are amplified using a barcoded primer, allowing up to 48 samples to be multiplexed and sequence variations interrogated. The NEXTflex Small RNA Sequencing Kit incorporates AIR™ Ligase, a patent pending, truncated T4 RNA Ligase, which significantly increases the efficiency with which small RNAs are tagged with adapter, giving greater sequence depth. The new protocol is designed to significantly reduce adapter dimer formation prevalent in other protocols.

Strand specificity is retained through the use of adapters that ligate to the 5’‐phosphate and 3’‐hydroxyl groups that most mature miRNAs possess. The kit can be used for mRNA directional sequencing. In combination with the NEXTflex™ Small RNA Barcode Primer kits, 48 samples can be profiled and studied simultaneously. Four sets of 12 NEXTflex™ Small RNA Barcode Primers, with embedded index sequences are available offering an improved multiplexing workflow and increased flexibility. This new automation-friendly format enables multiplexing of up to 384 samples. The ability to pool samples in an efficient way significantly decreases hands on time while providing robust data quality. The primer barcoding system utilizes a 6 nt index to differentiate up to 48 different samples on a single flow cell lane. In addition to small RNA sequencing, the NEXTflex Small RNA Sequencing Kit is compatible with mRNA directional, RIP-Seq and CLIP-Seq protocols.

The NEXTflex Small RNA Sequencing Kit and NEXTflex Small RNA Barcode Primers are the latest additions to Bioo Scientific’s portfolio of next generation sequencing solutions. These kits demonstrate Bioo Scientific’s commitment to developing innovative solutions to improve, simplify and reduce the costs of next generation sequencing.

About Bioo Scientific Corporation
Bioo Scientific Corporation is an Austin, TX based biotechnology company that provides innovative solutions to the life science industry. Bioo Scientific offers a complete portfolio of products that increase sensitivity, flexibility and speed to next-generation sequencing.

Bioo Scientific’s products are used around the world and can be purchased both directly or through our global distribution network with markets in more than 40 countries. For more information about Bioo Scientific’s NGS product line, visit www.biooscientific.com.

[monad]

In addition to small RNA sequencing, the NEXTflex Small RNA Sequencing Kit is compatible with mRNA directional, RIP-Seq and CLIP-Seq protocols.

I would appreciate to learn about the availability of strand strand specific (directional mRNA) kit from your company and whether that is compatible with Illumina TruSeq sequencing. Thanks!

[Bioo Scientific]

Hi Monad,

The NEXTflex™ Small RNA Sequencing Kit and the NEXTflex™ Small RNA Barcode Primers are compatible with strand specific directional mRNA Seq and the TruSeq cluster kits.

If you PM me your email address I can keep you updated about kits we will be launching soon.

[monad]

Excellent!

[magarine]

How much is the minimum amount of total RNA?

Hi, I wonder the minimum amount of total RNA to use your small RNA library kit.

And, if I want to start with small RNA as initial source, what amount of small RNA is proper?

[monad]

1 ug of input usually good for the Illumina prep. Do not know about the Nextflex.

[Bioo Scientific]

Hi Magarine and Monad,

You can start with as little as 500 ng - 1 ug of total RNA with the NEXTflex Small RNA Sequencing kit. If starting with enriched small RNA, we recommend using a quantity isolated from 1 - 10 ug of total RNA.

[volks]

hi, are the NEXTflex Small RNA Barcode Primers compatible with the Illumina TruSeq Small RNA adapters?

[Bioo Scientific]

Hi Volks,

Yes. The NEXTflex Small RNA Barcode Primers are compatible with the Illumina TruSeq Small RNA adapters and can be used with the NEXTflex Small RNA Seq Kit.

[danwiththeplan]

Strand specificity

Hi, just to clarify a question about strand specficity.

In Illumina Truseq system, the + strand is digested and the - strand is retained, and that's how strand specificity is maintained, such that only the - strand is sequenced (since the + strand is digested).

So for example, when running htseq-count you would count using the -reverse option (rather than the -no or -yes option) which says that htseq-count expects that the reverse strand is the only one sequenced.

In this system, which strand is retained? or, if this isn't how it works, how do you distinguish the strands ?

[Bioo Scientific]

Hi Dan,

Small RNA sequencing is a little different in that it involves the ligation of single stranded adapters directly to RNA molecules, rather than conversion to dsDNA and ligation of Y-shaped adapters, as is used in other RNA-Seq kits, such as the TruSeq kits and the NEXTflex Rapid Directional RNA-Seq Kits. Because of this, the sequence reported is that of the RNA molecule, or the + strand. Consequently direct ligation also allows retention of the 5' and 3' most bases of the RNA molecule, which typically isn't possible with dsDNA ligation strategy, as some of that information is lost during first and second strand synthesis.