Invitrogen™ Poly(A) Tail-Length Assay Kit

Description

• Poly adenylation of eukaryotic mRNAs plays a critical role in RNA metabolism, including stability, enhanced translation, nuclear export, and miRNA mediated gene regulation
• The Poly(A) Tail-Length Assay Kit allows you to quickly measure the poly(A) tails of multiple mRNAs in only three hours
• This kit utilizes a novel G/I extension technique that preserves the 3' end of RNAs and ensures accurate measurement of poly(A) tail-length
• And all reaction components have been optimized so you get quality data quickly
• Features Measure the poly(A) tail of multiple RNAs in under 3 hrs using common lab equipment Accurate measurement of poly(A) tail length compared to other assays One kit includes all necessary reagents for 5 G/I tailing, 20 RT, and 80 PCR reactions Technology The Poly(A) Tail-Length Assay Kit uses four key steps to enable poly(A) tail-length determination (see figure)
• Step 1—poly(A) polymerase adds a limited number of guanosine and inosine residues to the 3'-ends of poly(A)-containing RNAs
• Step 2—the tailed-RNAs are converted to DNA through reverse transcription using the newly added G/I tails as the priming sites
• Step 3—PCR amplification products are generated using two primer sets
• A gene-specific forward and reverse primer set designed upstream of the polyadenylation site (e.g., the 3'-UTR) is produced as a control for the gene-of-interest
• The second set of primers uses the gene-specific forward primer and the universal reverse primer provided with the kit to generate a product that includes the poly(A) tails of the gene-of-interest
• Step 4—the PCR products are separated on an agarose or polyacrylamide gel (see figure)
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