rna purification

[renuka22]

Hello,

I have RNA concentration around 15-18 ng/ul (260/280 is 1.5-1.8 but 260/230 is very low ~0.5) on which I need to do polyA library preparation and sequencing. The company said you need to purify the samples. If I purify the samples, I will lose more RNA. Is it risky to go ahead with library preparation without purifying the samples?

Thanks

[huguesparri]

Hello,

Did you only use the nanodrop to quantify your RNA samples or did you also perform some integrity checking on bioanalyzer?
Your concentration is pretty low and your nanodrop ratios are not good: you may have less material than you think...
You will find attached to this message a bulletin from Nanodrop explaining all this.

If you plan to purify your sample, you may try Agencourt RNA cleanXP beads from Beckman Coulter.

Is it risky to go ahead with library preparation without purifying the samples?

I will answer this question using 2 other questions:
- Do you think the company asked you to purify your samples just to ruin your day?
- Do you think performing molecular biology on contaminated samples is risky?

Regards,

Hugues