Small RNA (NOT microRNA) -seq

[SeqByChoice]

Hi !
Has anybody had any experience sequencing small RNA libraries?
meaning: 40-200nt RNAs
Any hints on good library prep protocols would be great!
As would any ideas on required sequencing depth for mammlian libraries?

[HESmith]

The most important consideration is to start with high-quality RNA (to avoid degradation products). You'll need a method to remove ribosomal RNA (5S is ~120bp) before size selection. If your RNAs of interest are not polyadenylated, then you'll also need to use a RT protocol that doesn't rely on oligo-dT priming.

[Biochain Rep]

If your RNA is fragmented, I wonder if something like RiboZero could at least reduce the amount of rRNA sequence generated. Also, if you're still test6ing kits, we at Biochain have one (www.biochain.com).