I am planning to deep sequence material from 13 samples. For this, I am isolating RNA from various organs, reverse transcribing with a gene specific primer and performing a multiplex PCR (all products are ~400 bp). Then, I will ligate a barcode to the PCR product from each sample and sequence. Since I am jumping into the middle of the standard 454 prep, can someone please tell me how much starting material I need for each sample for a full 454 plate?
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