Ultra low stranded mRNA Seq SMARTer kit:

Preparation of RNA samples for sequencing on the Illumina HiSeq. Described here is a library preparation service only. Once the library is prepared, you will also need to request (and pay for) the sequencing of that library.

Ribosomal RNA depleted RNA is input into the SMARTer stranded workflow and made into cDNA.

The first strand cDNA is purified and the library is amplified by PCR with the Illumina Indexing Primer set. The resultant library is then PCR amplified to have enough material for the sequencer.

 

Ultra low stranded mRNA Seq SMARTer kit:

Platform
Illumina HiSeq Libraries
Category
Gene Expression Profiling
Primary Contact
Typical Cost Click here to view internal pricing list!
Click here to view external pricing list!
Typical Output

The SMARTer stranded kit will yield an Illumina ready to sequence library with an insert size of 180bp 20 ul of ~10nM.

How to Request

Submit samples here: http://seqcore.brcf.med.umich.edu/cgi-bin/Solexa/login.cgi

Please deliver RNA to NCRC. Courier service from other sites will not guarantee proper temp is maintained!

Sample Requirements

The total fraction of 100pg – 100ng full length, intact total RNA that has been depleted of ribosomal RNA.

This kit has a separate protocol to deal with degraded RNA Samples, such as FFPE RNA which decreases the fragmentation time.

Typical Turnaround

3 weeks.

Critical Information

Requires ribosomal removal:

 

Ribo Gone requires 10-100ng total RNA abd us the ribosomal removal kit for human mouse and rat available by Clontech:

http://www.clontech.com/US/Products/cDNA_Synthesis_and_Library_Construction/Next_Gen_Sequencing_Kits/Ribosomal_RNA_Removal

 

The SMARTer stranded protocol has been optimized for cDNA synthesis starting from 10pg of total RNA. However if you RNA sample is not limiting, Clontech recommend that you start with more total RNA (up to 10 ng). Purified total RNA should be in nuclease free water.