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FORMAT 
Sure-RACE cDNA 5 End Discovery Panels are assembled from 24 major human tissues or 24 major mouse tissues at different developmental stages. The RACE-ready cDNAs are provided in two concentrations (5x and 1x) in 48-well (6 x 8 wells) PCR plates to accommodate the analysis of both rare and abundant transcripts. Four panels are provided per order to allow replication of the findings and assurance of the specificity of the observed RACE products.
Kit Components
- Four identical 48-well PCR plates containing two dilutions of 24 human or 24 mouse RACE-ready cDNAs
- Adaptor primers for PCR
- Control primers for transferrin receptor
- Quanti-Ladder DNA Marker
Analysis is performed using two rounds of PCR. The first round uses Adaptor Primer 1 and a gene-specific primer to enrich for the specific cDNAs. The second round uses Adaptor Primer 2 and a nested gene-specific primer to further amplify the specific target(s). Approximately 20 PCR cycles are routinely performed in the first round, followed by 30-35 cycles in the second round (nested) PCR. The specificity of the gene-specific primers is a critical factor.
By performing a nested PCR analysis, using the two contiguous adaptor-specific primers provided with the kit and two contiguous gene-specific primers designed by the investigator, Sure-RACE panels [1] markedly increase the probability of completing the 5' ends of transcripts which are rare or have complex secondary structures; [2] allow the identification of alternate RNA start-sites either within the same tissue, in different tissues or at different developmental stages; and [3] provide evidence for alternate RNA splicings at the 5' end, such as the use of alternate first exons or an identical first exon but alternate downstream exons, which may result in proteins with nonidentical amino acid sequences
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