Stranded mRNA Seq
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.
Total RNA is input into the workflow where mRNA is isolated using oligo d(T), cDNA is made with reverse transcriptase. Second strand synthesis incorporates uracils in place of thymines. The cDNA is end repaired A tailed and Illumina adapters are ligated. The resultant library is then PCR amplified to have enough material for the sequencer. The enzyme in the PCR reaction does not recognize the uracils and therefore does not amplify the 2nd strand.
|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||~30 ul of 10nM prepared library with a 120 bp insert, you will also get TapeStation reports of the quality of your input RNA and the resultant library.|
|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||100ng -4ug of good quality, RINs of 8 or higher, DNase treated total RNA in 25-50ul|
|Critical Information||RNA must be delivered on dry ice to NCRC and cannot be left at our pick up sites on campus.|
|Additional Information||Please discuss your RNA Seq project with your bioinformaticist or contact the University of Michigan's Bioinformatics Core (firstname.lastname@example.org) before you begin your project.
For kit information please see Illumina's site:
Due to limited space in the cores freezer's we can only keep samples for 3 months. If you would like your samples returned to you after sequencing please contact the core.