Characterization of genomically amplified segments using PCR: Optimizing relative-PCR for reliable and simple gene expression and gene copy analyses

Molecular cloning and characterization of a granulin-containing cysteine protease SPCP3 from sweet potato (Ipomoea batatas) senescent leaves

“…The 5 0 (Y508A: TGTGCTGACGATGATTCG CTCCGTC) and 3 0 (Y508B: ACTGCAAATTATGCACT-GAATCCTC) primers were used to amplify the fulllength, double-stranded cDNAs for semi-quantitative analysis according to the method of Jonson et al (2000). The full-length cDNA of SPCP3 in the recombinant plasmid was labeled with digoxigenin-11-dUTP nucleotides as the probe to detect semiquantitative RT-PCR products using DNA gel blot hybridization and CSPD substrate (Boehringer Mannheim) as described previously.…”

“…The temporal and spatial expression patterns of SPCP3 were studied with semi-quantitative RT-PCT according to Jonson et al (2000). The amount of PCR-amplified product was detected by ethidium bromide (EtBr) staining and Dig-labeled probe hybridization.…”

Molecular cloning and characterization of a granulin-containing cysteine protease SPCP3 from sweet potato (Ipomoea batatas) senescent leaves

“…The 5 0 (Y508A: TGTGCTGACGATGATTCG CTCCGTC) and 3 0 (Y508B: ACTGCAAATTATGCACT-GAATCCTC) primers were used to amplify the fulllength, double-stranded cDNAs for semi-quantitative analysis according to the method of Jonson et al (2000). The full-length cDNA of SPCP3 in the recombinant plasmid was labeled with digoxigenin-11-dUTP nucleotides as the probe to detect semiquantitative RT-PCR products using DNA gel blot hybridization and CSPD substrate (Boehringer Mannheim) as described previously.…”

“…The temporal and spatial expression patterns of SPCP3 were studied with semi-quantitative RT-PCT according to Jonson et al (2000). The amount of PCR-amplified product was detected by ethidium bromide (EtBr) staining and Dig-labeled probe hybridization.…”

Molecular cloning and characterization of a granulin-containing cysteine protease SPCP3 from sweet potato (Ipomoea batatas) senescent leaves

“…The 5 0 (Y508A: TGTGCTGACGATGATTCG CTCCGTC) and 3 0 (Y508B: ACTGCAAATTATGCACT-GAATCCTC) primers were used to amplify the fulllength, double-stranded cDNAs for semi-quantitative analysis according to the method of Jonson et al (2000). The full-length cDNA of SPCP3 in the recombinant plasmid was labeled with digoxigenin-11-dUTP nucleotides as the probe to detect semiquantitative RT-PCR products using DNA gel blot hybridization and CSPD substrate (Boehringer Mannheim) as described previously.…”

“…The temporal and spatial expression patterns of SPCP3 were studied with semi-quantitative RT-PCT according to Jonson et al (2000). The amount of PCR-amplified product was detected by ethidium bromide (EtBr) staining and Dig-labeled probe hybridization.…”

Molecular cloning and characterization of a granulin-containing cysteine protease SPCP3 from sweet potato (Ipomoea batatas) senescent leaves

“…The 5 0 (Y508A: TGTGCTGACGATGATTCG CTCCGTC) and 3 0 (Y508B: ACTGCAAATTATGCACT-GAATCCTC) primers were used to amplify the fulllength, double-stranded cDNAs for semi-quantitative analysis according to the method of Jonson et al (2000). The full-length cDNA of SPCP3 in the recombinant plasmid was labeled with digoxigenin-11-dUTP nucleotides as the probe to detect semiquantitative RT-PCR products using DNA gel blot hybridization and CSPD substrate (Boehringer Mannheim) as described previously.…”

“…The temporal and spatial expression patterns of SPCP3 were studied with semi-quantitative RT-PCT according to Jonson et al (2000). The amount of PCR-amplified product was detected by ethidium bromide (EtBr) staining and Dig-labeled probe hybridization.…”