Homemade Illumina mRNA-SEQ protocols

[kbushley]

I'm new to mRNA-SEQ and wondering if anyone has a good homemade protocol that mimicks Illumina's standard mRNA-SEQ protocol. In particular wondering about what fragmentation methods seems to work best and if Illumina uses proprietary random hexamer primers or if Invitrogen random primers seem to work just as well.


Thanks!

[jgibbons1]

I can admittedly take this chance to self advertise:

Gibbons JG, Janson EM, Hittinger CT, Johnston M, Abbot P, Rokas A: Benchmarking next-generation transcriptome sequencing for functional and evolutionary genomics. Mol Biol Evol 2009, 26(12):2731-2744.

We reports a full mRNA-SEQ protocol in the methods section. Please let me know if you have any questions. We used a zinc acetate shearing for fragmentation.

[kbushley]

Hi John,

Nothing wrong with a little self promotion -...especially since it's an interesting paper...thanks! I think I will give this a shot.