Invitrogen generacer kit manual

The GeneRacer™ Kit provides a method to obtain full-length 5' and 3' ends of cDNA using known cDNA sequence from expressed sequence tags (ESTs), subtracted cDNA, differential display, or library screening. The kit ensures the amplification of only full-length transcripts via elimination of truncated messages from the amplification process. Each GeneRacer Kit contains the GeneRacer box, an RT box, S.N.A.P. columns, and a TOPO Cloning Kit. The GeneRacer and the RT box contain sufficient reagents for five cDNA reactions plus one control reaction and primers for 50 PCR reactions. Included in the GeneRacer box are the enzymes CIP, TAP, T4 RNA ligase, and their buffers, the GeneRacer Format: Kit. The 5′ ends of RNAs were amplified with the GeneRacer kit (Invitrogen Ltd.), which is designed to amplify only 5′ ends of m7 GpppN-capped RNA molecules Briefly, the procedure involves (i) dephosphorylation of the 5′ ends of noncapped RNAs with calf intestinal phosphatase, (ii) removal of the cap structure from the capped RNAs with tobacco acid pyrophosphatase, thereby .

MATERIALS AND METHODS Identification and verification of the 5′ and 3′ end of the RAGE mRNA. To enable the design of primers to screen full-length RAGE mRNAs for novel splice forms, the start (5′ end) and finish (3′ end) of the human RAGE mRNA was determined by RNA-ligase mediated-RACE (RLM-RACE), using the GeneRacer kit (Invitrogen, Carlsbad, CA, USA). Invitrogen champion pet directional topo expression kit with bl21 star de3 one shot chemically competent e. 1 methods overview introduction topo ta cloning * provides a highly efficient, 5 minute, one-step cloning strategy topo cloning for the direct insertion of taq polymerase-amplified pcr products into a plasmid vector. (Invitrogen) and random primers. For RACE, 5mg of total RNA were reverse transcribed into cDNA using Superscript reverse transcriptase III (Invitrogen) and poly(dT) primers provided in the GeneRacer kit. Both RACE and RACE cDNAs were amplified by PCR. The primers for PCR amplification are listed in Table 1.

Generacer manual 1. Instruction Manual GeneRacer™ Kit For full-length, RNA ligase-mediated rapid amplification of 5´ and 3´ cDNA ends (RLM-RACE) Catalog nos. L; L; L; L Version L 8 April 2. ii 3. The GeneRacer™ Kit provides a method to obtain full-length 5' and 3' ends of cDNA using known cDNA sequence from expressed sequence tags (ESTs), subtracted cDNA, differential display, or library screening. The kit ensures the amplification of only full-length transcripts via elimination of truncated messages from the amplification process. GeneRacer™ Kit with SuperScript™ III RT and TOPO TA Cloning™ Kit for Sequencing Invitrogen™ The GeneRacer™ Kit provides a method to obtain full-length 5' and 3' ends of cDNA using known cDNA sequence from expressed sequence tags (ESTs), subtracted cDNA, differential display, or library screening.