New RNA-Seq library prep kit

[epibio]

Epicentre Biotechnologies announces the availability of the new ScriptSeq(TM) mRNA-Seq Library Preparation Kit (Illumina-compatible). The kit uses a unique, patented terminal-tagging technology that simplifies strand-specific tagging and the preparation of directional mRNA-Seq libraries from rRNA-depleted or poly(A) RNA.

Starting from just 50 ng of rRNA-depleted mRNA, or poly(A) RNA, the ScriptSeq kit produces adaptor-tagged libraries in about 3 hours without the need for end-repair, adaptor ligations, shearing the cDNA, or gel electrophoresis.

For more information, please see the product page.

[mnkyboy]

Do you have any information on how this would perform for whole transcriptome analysis and not just mRNA-seq?

[epibio]

The kit is actually suitable for whole-transcriptome analysis, since it uses random-primed cDNA synthesis. The choice of mRNA-Seq or WT would be determined by whether you were using poly(A)-enriched or rRNA-depleted RNA, respectively.

[cldashie]

Hi - I was wondering if your adapters were paired-end ready for the mRNA seq kit, and if your adapters are computable with Illumina's indexes and multiplexing pcr primers.

[epibio]

Yes, the adaptors are designed to enable paired-end reads. They are also suitable for multiplexing, either with our barcodes kit or with barcodes that you design.

[upendra_35]

do you know when will the Ribo-zero rRNA kit will be available for plant RNA?

[epibio]

Answered via PM.

[MaizeMaze]

Can you PM me the ribo-zero for plant info too?
Thanks!

[epibio]

I don't think I can PM you as yet, since you're a new user. However, we expect the plant kits to be released early next year. If you're interested in beta-testing the kits, please contact me using the e-mail link in my profile.

[arabidopsis]

ScriptSeq kit ia supposed to produce adaptor-tagged libraries from as little as 50 ng of rRNA-depleted mRNA. My question is addressed to the the vendor (epibio?) as well as to the experienced users.
I produced some libraries using 200 ng rRNA-depleted RNA as a starting material. After 10 cycles of PCR amplification, recommended by Epicentre, DNA is barely detectable by spectrophotometry (concentration less than 50 ng/ul) What is the expected yield of double-stranded DNA (final product)? Is it normal to have such low yield, or something have gone wrong, or else my spec is just out of order?

[HESmith]

50 ng/ul is plenty! One ng of library is sufficient for an entire flow cell (~500 million reads on a HiSeq).

[arabidopsis]

Thanks HESmith!
I have no experience with those libraries; neither do other people in the lab. This would be our first RNAseq project , thus I have been quite nervous about it .

rRNA removal kit of Epicentre worked pretty good, by the way. qPCR shown 78,000-fold reduction of rRNA amplicon signal.

[epibio]

@ arabidopsis: Assuming you had 50 ul after PCR, that should be plenty, as HESmith said. For the kit QC, we use 100 ng of poly(A)+ RNA as input, and obtain around 150 ng of the cDNA library.

[RCJK]

@epibio: have you had any feedback from anyone using the mRNA-seq kits for 454? Are you able to point me to anywhere that would have this info? Thanks.

[epibio]

We have not heard from any customers using the ScriptSeq Titanium-compatible kit. Most of our development efforts have focused on the Illumina platform, but the Titanium-compatible kit is very similar to the Illumina-compatible one, with the exception of adaptor sequences, and should perform equivalently.