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.
|Platform||Illumina HiSeq 2500, HiSeq 4000, NextSeq, and MiSeq.
|Category||Gene Expression Profiling|
|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||Sample Submission: login through https://client-seqcore.brcf.med.umich.edu/ then choose ‘Illumina Sequencing’
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.
|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:
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.
|Additional Information||Due to limited space in the cores freezer's we can only keep samples for 6 months. If you would like your samples returned to you after sequencing please contact the core.