TruSeq small RNA final products Peaks

[shahideh]

Hi everyone,

I have tried to make small RNA libraries from insect RNAs and I should say that it was my first experience. I used TruSeq small RNA kit. I have attached my final products (samples 1-3, 4 is just water) run in Experion DNA 1K chip from Bio-Rad after gel purification and ethanol precipitation. I appreciate if you guys let me know if you think they look good to send for sequencing.
My ultimate goal is discovering known and novel viruses in insect. So, I am looking for any kind of small RNAs (18-30 nt).

Thanks

[kcchan]

Your libraries look perfect! The goal of the gel cut is to remove the primer and adapter dimer bands at ~125 and 135bp while keeping your small RNA libraries, which in your case are ~153-165bp. Looking from these traces you have nothing but the library peak. The yields also look good enough for pooling and sequencing.

[shahideh]

Thanks so much for your reply

[F2000]

Hi,
Which method did you use for isolating the small RNA fragment from the adapter dimer.

Thanks

[shahideh]

HI F2000,

I did gel purification. I followed the TruSeq protocol