Does anyone know how to duplicate the 1-step elution/priming/fragmentation step from the TruSeq mRNA sequencing protocol using homemade reagents? I have oligo(dT) beads and buffers, fragmentation buffer (Zn-based, from Ambion), and random primers. My problem is that I don't know how to mix the fragmentation buffer and random primers to make the reagent that elutes and fragments while still being compatible with downstream cDNA synthesis. Do I need to use a different fragmentation reagent?
Anyone who has used the Illumina protocol knows that it saves a good bit of time - and allows 96-well plate processing - not to have to purify the RNA out of the fragmentation buffer before proceeding with RT.
Thanks!
Anyone who has used the Illumina protocol knows that it saves a good bit of time - and allows 96-well plate processing - not to have to purify the RNA out of the fragmentation buffer before proceeding with RT.
Thanks!
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