341 F CCTACACGACGCTCTTCCGATCTN (barcode) CCTACGGGNGGCWGCAG, reverse primers: 805 R GACTGGAGTTCCTTGGCACCCGAGAATTCCAGACTACHVGGGTATCTAATCC). The reaction mixtures (50 µl) contained 5 µl of 10 × PCR reaction buffer (TakaRa, Japan), 10 ng of DNA template, 0.5 µl of each primer, 0.5 µl of dNTPs and 0.5 µl of Platinum Taq DNA polymerase (TakaRa, Japan). The PCR conditions were as follows: 94 °C for 3 min, 94 °C for 30 s, annealing at 45 °C for 20 s and 65 °C for 30 s, which was repeated for 5 cycles, followed by 94 °C for 20 s, 55 °C for 20 s and 72 °C for 30 s, which was repeated for 20 cycles, before a final elongation at 72 °C for 5 min. The PCR product was excised from the 1.5% agarose gel and purified using a QIAquick Gel Extraction Kit.
Platinum taq dna polymerase
Platinum Taq DNA polymerase is a thermostable DNA polymerase enzyme used for PCR amplification of DNA. It catalyzes the synthesis of new DNA strands complementary to a DNA template.
Lab products found in correlation
6 protocols using platinum taq dna polymerase
Soil DNA Extraction and 16S rRNA Amplification
341 F CCTACACGACGCTCTTCCGATCTN (barcode) CCTACGGGNGGCWGCAG, reverse primers: 805 R GACTGGAGTTCCTTGGCACCCGAGAATTCCAGACTACHVGGGTATCTAATCC). The reaction mixtures (50 µl) contained 5 µl of 10 × PCR reaction buffer (TakaRa, Japan), 10 ng of DNA template, 0.5 µl of each primer, 0.5 µl of dNTPs and 0.5 µl of Platinum Taq DNA polymerase (TakaRa, Japan). The PCR conditions were as follows: 94 °C for 3 min, 94 °C for 30 s, annealing at 45 °C for 20 s and 65 °C for 30 s, which was repeated for 5 cycles, followed by 94 °C for 20 s, 55 °C for 20 s and 72 °C for 30 s, which was repeated for 20 cycles, before a final elongation at 72 °C for 5 min. The PCR product was excised from the 1.5% agarose gel and purified using a QIAquick Gel Extraction Kit.
Nrf2 Regulation: Chemical and Molecular Insights
Blood Genomic DNA Extraction and Analysis
Mosquito Species Identification by PCR
[44 (link),45 (link)]. Amplification was performed in a 25 μL reaction containing 2 μL of template DNA, 2.5 μL of 10 × PCR buffer, 0.75 μL of 50 mM MgCl2, 2 μL of 2.5 mM of each dNTP, 0.5 μL of 10 μM of each primer, and 0.625unit Platinum®Taq DNA Polymerase (TaKaRa, China). The cycling conditions were as follows: initial denaturation at 94°C for 2 min, 30 cycles of 30s denaturation at 94°C, 30s annealing at 48°C and 30s extension at 72°C followed by a final extension of 10 min at 72°C. Amplification products were examined on a 2.5% agarose gel electrophoresis. The species was determined by the size of the PCR product (1,077 bp for An. sinensis and 604 bp for An. vagus). Molecular identification was conducted for all mosquitoes subjected to insecticide resistance bioassay.
Quantifying Osteogenic Marker Expression
Primers for RUNX2, osteopontin, DLX5, osterix, and osteocalcin were used as follows: RUNX2: sense 5′-GATGACACTGCCACCTCTGA-3′, antisense 5′-CAGCGTCAACACCATCATTC-3′; osteopontin: sense 5′-TCTGATGAGACCGTCACTGC-3′, antisense 5′-TGTCCTTGTGGCTGTGAAAC-3′; DLX5: sense 5′-TCTCTAGGACTGACGCAAACA-3′, antisense 5′-GTTACACGCCATAGGGTCGC-3′; ssterix: sense 5′-GATAGTGGAGACCTTGCTCGTAG-3′, antisense 5′-GAGGTCACAGGGTATGAGAAGAG-3′; osteocalcin: sense 5′-AGGACCATCTTTCTGCTCACTC, antisense 5′-CTGCCAGAGTTTGGCTTTAG.
Soil DNA Extraction and 16S rRNA Amplification
were used in the amplification of the V3-V4 hypervariable regions of 16S rRNA via PCR from the microbial genomic DNA. The PCR reaction mixture (50 μl) comprised of 5 μl of 10 × PCR reaction buffer (TakaRa, Japan), 10 ng of DNA template, 0.5 μl of each primer, 0.5 μl of dNTPs and 0.5 μl of Platinum Taq DNA polymerase (TakaRa, Japan). Thermocycler amplification conditions were set at: initial 94ºC for 3 min, 94ºC for 30 s, annealing at 45ºC for 20 s and 65ºC for 30 s, which was repeated for 5 cycles, followed by 94ºC for 20 s, 55ºC for 20 s and 72ºC for 30 s, which was repeated for 20 cycles, before a final elongation at 72ºC for 5 min. Specificity assessment of the PCR amplification product was assessed on 1.5% agarose gel and thereafter purified using a QIAquick Gel Extraction Kit.
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