We have been having issues with our RNA from FFPE when submitting for sequencing. We are using the Illumina kit RS-122-2201, TruSeq Stranded Total RNA RboZroHMN/Mse/Rat with ribo depletion.
Our samples are degraded so we have decided to use either no fragmentation or minimal fragmentation (Covaris) to start with.
We are also considering using up to 750 ng of input RNA.
I'm concerned that we may be using too much to start with and that can be problematic. We have a CORE facility that makes our libraries, so I'm not 100% familiar with the protocol for library construction - just a rough idea.
If I don't need to use this much material, I would love to be able to save some for other downstream testing.
Thank you.
Our samples are degraded so we have decided to use either no fragmentation or minimal fragmentation (Covaris) to start with.
We are also considering using up to 750 ng of input RNA.
I'm concerned that we may be using too much to start with and that can be problematic. We have a CORE facility that makes our libraries, so I'm not 100% familiar with the protocol for library construction - just a rough idea.
If I don't need to use this much material, I would love to be able to save some for other downstream testing.
Thank you.
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