Selected article for: "primer mix and total volume"
Author: Alves, Christian D.B.T.; Budaszewski, Renata F.; Torikachvili, Marcela; Streck, André F.; Weber, Matheus N.; Cibulski, Samuel P.; Ravazzolo, Ana P.; Lunge, Vagner R.; Canal, Cláudio W.
Title: Detection and genetic characterization of Mamastrovirus 5 from Brazilian dogs
  • Document date: 2018_2_2
    • ID: 29wagjw8_7
    Snippet: The cDNA was synthesized using SuperScript ® III Reverse Transcriptase Kit (Life Technologies, USA) using the reverse primers in a total volume of 20 L, following the manufacturer's instructions. The cDNA amplification was conducted in a final volume of 25 L containing 1× PCR buffer, 1.5 mM of MgCl 2 , 0.2 mM of dNTP mix, 0.2 M of each primer and 1 unit of Platinum ® Taq DNA Polymerase (Life Technologies, USA). The first round of RT-PCR screen.....
    Document: The cDNA was synthesized using SuperScript ® III Reverse Transcriptase Kit (Life Technologies, USA) using the reverse primers in a total volume of 20 L, following the manufacturer's instructions. The cDNA amplification was conducted in a final volume of 25 L containing 1× PCR buffer, 1.5 mM of MgCl 2 , 0.2 mM of dNTP mix, 0.2 M of each primer and 1 unit of Platinum ® Taq DNA Polymerase (Life Technologies, USA). The first round of RT-PCR screening was carried out with an initial incubation at 94 • C for 3 min, 30 cycles of amplification consisting of denaturation at 94 • C for 1 min, annealing at 50 • C for 1 min, and extension at 72 • C for 1 min. The second round was performed in a final volume of 25 L that contained 2 L of the first reaction product and the thermocycler conditions were the same as those used for the first round. The MAstV5-specific RT-PCR with specific and internal control primers was performed as a multiplex protocol. Cycling conditions were an initial cycle at 94 • C for 5 min, 25 cycles of denaturation at 94 • C for 30 s, annealing at 58 • C for 30 s and polymerization at 72 • C for 1 min, which was followed by a final extension cycle at 72 • C for 7 min. To confirm the specific amplification of MAstV5, RT-PCR products

    Search related documents:
    Co phrase search for related documents
    • cdna amplification and reaction product: 1
    • cdna amplification and reverse primer: 1, 2, 3, 4
    • cdna amplification and RT PCR product: 1, 2
    • dNTP mix and extension cycle: 1
    • dNTP mix and PCR buffer: 1, 2
    • dNTP mix and reverse primer: 1
    • extension cycle and initial cycle: 1
    • extension cycle and PCR buffer: 1
    • extension cycle and reverse primer: 1, 2
    • final volume and PCR buffer: 1
    • final volume and reaction product: 1, 2
    • final volume and reverse primer: 1, 2, 3, 4, 5, 6, 7
    • internal specific control and RT PCR screening: 1
    • PCR buffer and reverse primer: 1, 2, 3
    • reaction product and reverse primer: 1
    • reaction product and RT PCR product: 1, 2