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Old 03-08-2013, 10:26 AM   #1
shahideh
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Default TruSeq small RNA final products Peaks

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
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File Type: pdf Finall library products.pdf (152.8 KB, 121 views)
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Old 03-10-2013, 10:03 PM   #2
kcchan
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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.

Last edited by kcchan; 03-10-2013 at 10:05 PM. Reason: Derp, didn't read the initial post fully. Sample 4 is NTC.
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Old 03-12-2013, 09:19 AM   #3
shahideh
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Thanks so much for your reply
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Old 03-14-2013, 09:02 AM   #4
F2000
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Hi,
Which method did you use for isolating the small RNA fragment from the adapter dimer.

Thanks
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Old 03-14-2013, 10:06 AM   #5
shahideh
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HI F2000,

I did gel purification. I followed the TruSeq protocol
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