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  • FAQ: What protocol do I use if starting with previously isolated mRNA or ribosomal depleted RNA?

    Reaction Setup for 10ng-100ng previously isolated mRNA or ribosomal depleted RNA:

    1.Purified mRNA or ribosomal depleted RNA(10-100ng) 5ul
    First Strand Synthesis Reaction Buffer 4ul
    Random Primers 1ul
    final Volume 10ul

    2. incubate the sample for 94C for 15 minutes (this is to achieve 200nt size fragments, refer to appendix A for larger sizes)

    3. Transfer tube to ice and proceed to First Strand synthesis in the appropriate chapter of the manual.