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[Music] thank you [Music] the overall goal of this procedure is to determine the sequences of mRNA transcripts from the three prime end to Regions within the protein coding region by using transcript specific nested primers in two subsequent pcrs and sequencing the purified PCR products this method can help answer key questions in the genetic field such as determining the polar dilation signal the stop codon and the sequence of the three prime untranslated region of a transcript the main advantage of this technique is that it can be applied to any transcript if the sequence of a small region of the open reading frame is known the implication of this technique extend toward cancer diagnosis because it can detect tumor-specific Fusion genes and splice variants we first had the idea for this method when we identified novel transcripts with previously uncharacterized three-prime untranslated regions to begin the protocol thaw the phenol and guanidine isothiocyanate cell mixture on ice onc
