Marathon-Ready cDNAs—cDNAs made from high-quality Premium Poly A+ RNA and ligated to the Marathon® Adaptor—are ready for 5'- and 3'-RACE PCR (1). Each Marathon-Ready cDNA is a premade, tissue-specific “pool” of double-stranded cDNA from which full-length genes can be amplified by using sets of gene-specific primers. They can also be used to study tissue-specific gene expression and to find polymorphic forms of mRNA or mRNA belonging to a multigene family

Marathon-Ready cDNA is synthesized from Premium RNA using a procedure optimized to produce full-length cDNA and to eliminate 3' heterogeneity (2, 3). After synthesis, blunt ends are created and the Marathon® Adaptor is ligated to both ends of the double-stranded cDNA.

Each Marathon-Ready cDNA is sufficient for 30 reactions (Conc. ~ 0.1 ng/μl) and includes an adaptor primer, nested adaptor primer, 5' and 3' G3PDH control primers, a User Manual (PT1156-1), and a Protocol-at-a-Glance (PT1156-2). To perform 5'- and 3'-RACE with Marathon-Ready cDNA, all you need is your gene specific primers and a polymerase mix for PCR. We recommend our Advantage® 2 Polymerase Mix.

Note: cDNA source may vary from lot to lot. For current source information, please refer to the Certificate of Analysis accompanying each Marathon-Ready cDNA.

Figure 1. Generation of full-length cDNA by end-to-end amplification from adaptor-ligated ds cDNA. RACE PCR was performed. The template for Lanes 1–3 was Marathon-Ready Human Placenta cDNA (Cat. No. 639311); the template for Lanes 4 & 5 was Marathon-Ready Human Skeletal Muscle cDNA (Cat. No. 639313). Lane 1: 5'-RACE product for TFR cDNA. Lane 2: 3'-RACE product for TFR cDNA. Lane 3: full-length TFR cDNA (5.1 kb) generated by end-to-end amplification using 5' and 3' TFR primers. Lane 4: nearlyfull-length IGFR2 cDNA (8.8 kb) amplified with 5' IGFR2 and 3' IGFR2 primers. Lane 5: full-length IGFR1 cDNA (5.0 kb) amplified with primers 5'-IGFR and 3'-IGFR. Lane M: 1-kb DNA size ladder.

 

Figure 2. Marathon® RACE products obtained using G3PDH primers to amplify Marathon-Ready Human, Mouse, and Rat Heart cDNAs. RACE PCR was performed using the AP1 primer and the positive control G3PDH primers. The templates used were Marathon-Ready Human Heart (Cat. No. 639304), Mouse Heart (Cat. No. 639404), and Rat Heart (Cat. No. 639416) cDNAs. Lane M: 1-kb DNA size ladder.