Amplification:Article Title:
Article Snippet: The purified PCR product was recycled and cloned to the pMD18-T vector (TaKaRa), and positive clones were selected for sequencing. .. Amplification of IL-6 3′ terminal sequence of S. chuatsi (1) Synthesis and Purification of the First Strand of cDNA Reverse transcription was performed on the total extracted RNA using SUPERSCRIPT II RT enzyme and the primer 3′CDS primer A (SMARTerTM RACE cDNA Amplification Kit, Clontech). .. The used primer was 3′CDS primer A, and the other components and conditions were the same as those of 5′ terminal amplification. (2) Quick Amplification of cDNA Ends The first round PCR amplification was performed using the primers GSP-IL-6-4 and UPM using the cDNA synthesized previously as a template, and the 3′-RACE reaction system was added in the following order (the reaction conditions were the same as those of the 5′ ends amplification): sterilized, distilled water 31.5 μl 10 × PCR buffer 5.0 μl 25 mM MgCl2 3.0 μl 10 mM dNTP mix 1.0 μl 10 μM GSP-IL-6-4 2.0 μl 10 uM UPM 2.0 μl dC-tailed cDNA 5.0 μl Taq DNA polymerase 0.5 μl final volume 50 μl The PCR product from the first round amplification was diluted 50 times to perform the second round PCR amplification, the other systems were the same as those of the first round PCR amplification other than the primers GSP-IL-6-5 and UPM, and the reaction conditions were the same as those of 5′ amplification.
Article Title:
Article Snippet: The purified PCR product was recycled and cloned to the pMD18-T vector (TaKaRa), and positive clones were selected for sequencing. .. Amplification of IL-6 3′ Terminal Sequence of S. chuatsi (1) Synthesis and Purification of the First Strand of cDNA Reverse transcription was performed on the total extracted RNA using SUPERSCRIPT II RT enzyme and the primer 3′CDS primer A (SMARTerTM RACE cDNA Amplification Kit, Clontech). ..
Sequencing:Article Title:
Article Snippet: The purified PCR product was recycled and cloned to the pMD18-T vector (TaKaRa), and positive clones were selected for sequencing. .. Amplification of IL-6 3′ terminal sequence of S. chuatsi (1) Synthesis and Purification of the First Strand of cDNA Reverse transcription was performed on the total extracted RNA using SUPERSCRIPT II RT enzyme and the primer 3′CDS primer A (SMARTerTM RACE cDNA Amplification Kit, Clontech). .. The used primer was 3′CDS primer A, and the other components and conditions were the same as those of 5′ terminal amplification. (2) Quick Amplification of cDNA Ends The first round PCR amplification was performed using the primers GSP-IL-6-4 and UPM using the cDNA synthesized previously as a template, and the 3′-RACE reaction system was added in the following order (the reaction conditions were the same as those of the 5′ ends amplification): sterilized, distilled water 31.5 μl 10 × PCR buffer 5.0 μl 25 mM MgCl2 3.0 μl 10 mM dNTP mix 1.0 μl 10 μM GSP-IL-6-4 2.0 μl 10 uM UPM 2.0 μl dC-tailed cDNA 5.0 μl Taq DNA polymerase 0.5 μl final volume 50 μl The PCR product from the first round amplification was diluted 50 times to perform the second round PCR amplification, the other systems were the same as those of the first round PCR amplification other than the primers GSP-IL-6-5 and UPM, and the reaction conditions were the same as those of 5′ amplification.
Article Title:
Article Snippet: The purified PCR product was recycled and cloned to the pMD18-T vector (TaKaRa), and positive clones were selected for sequencing. .. Amplification of IL-6 3′ Terminal Sequence of S. chuatsi (1) Synthesis and Purification of the First Strand of cDNA Reverse transcription was performed on the total extracted RNA using SUPERSCRIPT II RT enzyme and the primer 3′CDS primer A (SMARTerTM RACE cDNA Amplification Kit, Clontech). ..
Purification:Article Title:
Article Snippet: The purified PCR product was recycled and cloned to the pMD18-T vector (TaKaRa), and positive clones were selected for sequencing. .. Amplification of IL-6 3′ terminal sequence of S. chuatsi (1) Synthesis and Purification of the First Strand of cDNA Reverse transcription was performed on the total extracted RNA using SUPERSCRIPT II RT enzyme and the primer 3′CDS primer A (SMARTerTM RACE cDNA Amplification Kit, Clontech). .. The used primer was 3′CDS primer A, and the other components and conditions were the same as those of 5′ terminal amplification. (2) Quick Amplification of cDNA Ends The first round PCR amplification was performed using the primers GSP-IL-6-4 and UPM using the cDNA synthesized previously as a template, and the 3′-RACE reaction system was added in the following order (the reaction conditions were the same as those of the 5′ ends amplification): sterilized, distilled water 31.5 μl 10 × PCR buffer 5.0 μl 25 mM MgCl2 3.0 μl 10 mM dNTP mix 1.0 μl 10 μM GSP-IL-6-4 2.0 μl 10 uM UPM 2.0 μl dC-tailed cDNA 5.0 μl Taq DNA polymerase 0.5 μl final volume 50 μl The PCR product from the first round amplification was diluted 50 times to perform the second round PCR amplification, the other systems were the same as those of the first round PCR amplification other than the primers GSP-IL-6-5 and UPM, and the reaction conditions were the same as those of 5′ amplification.
Article Title:
Article Snippet: The purified PCR product was recycled and cloned to the pMD18-T vector (TaKaRa), and positive clones were selected for sequencing. .. Amplification of IL-6 3′ Terminal Sequence of S. chuatsi (1) Synthesis and Purification of the First Strand of cDNA Reverse transcription was performed on the total extracted RNA using SUPERSCRIPT II RT enzyme and the primer 3′CDS primer A (SMARTerTM RACE cDNA Amplification Kit, Clontech). ..
Reverse Transcription:Article Title:
Article Snippet: The purified PCR product was recycled and cloned to the pMD18-T vector (TaKaRa), and positive clones were selected for sequencing. .. Amplification of IL-6 3′ terminal sequence of S. chuatsi (1) Synthesis and Purification of the First Strand of cDNA Reverse transcription was performed on the total extracted RNA using SUPERSCRIPT II RT enzyme and the primer 3′CDS primer A (SMARTerTM RACE cDNA Amplification Kit, Clontech). .. The used primer was 3′CDS primer A, and the other components and conditions were the same as those of 5′ terminal amplification. (2) Quick Amplification of cDNA Ends The first round PCR amplification was performed using the primers GSP-IL-6-4 and UPM using the cDNA synthesized previously as a template, and the 3′-RACE reaction system was added in the following order (the reaction conditions were the same as those of the 5′ ends amplification): sterilized, distilled water 31.5 μl 10 × PCR buffer 5.0 μl 25 mM MgCl2 3.0 μl 10 mM dNTP mix 1.0 μl 10 μM GSP-IL-6-4 2.0 μl 10 uM UPM 2.0 μl dC-tailed cDNA 5.0 μl Taq DNA polymerase 0.5 μl final volume 50 μl The PCR product from the first round amplification was diluted 50 times to perform the second round PCR amplification, the other systems were the same as those of the first round PCR amplification other than the primers GSP-IL-6-5 and UPM, and the reaction conditions were the same as those of 5′ amplification.
Article Title:
Article Snippet: The purified PCR product was recycled and cloned to the pMD18-T vector (TaKaRa), and positive clones were selected for sequencing. .. Amplification of IL-6 3′ Terminal Sequence of S. chuatsi (1) Synthesis and Purification of the First Strand of cDNA Reverse transcription was performed on the total extracted RNA using SUPERSCRIPT II RT enzyme and the primer 3′CDS primer A (SMARTerTM RACE cDNA Amplification Kit, Clontech). ..
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