Bovine CD14 Promoter Amplification
Corresponding Organization : Rochester Institute of Technology
Other organizations : Ladoke Akintola University of Technology
Variable analysis
- Primer pairs used to amplify the ~1.6 kB promoter region of bovine CD14 gene
- Concentration of extracted genomic DNA quantified with a spectrometer
- Amplification of the ~1.6 kB promoter region of bovine CD14 gene
- Sequence of the amplified PCR products
- Volume of whole blood used for DNA extraction (200 μL)
- DNA extraction kit (Isolate II Genomic DNA extraction kit)
- Elution volume of extracted DNA (100 μL)
- Storage temperature of extracted DNA (4 °C)
- PCR master mix (EconoTaq Plus Green 2X PCR master mix)
- Volume of genomic DNA used for PCR (1 μL)
- Final PCR reaction volume (25 μL)
- PCR conditions (94 °C for 2 min, 35 cycles of 94 °C for 30 s, 59 °C for 30 s, 72 °C for 50 s, and final extension at 72 °C for 5 min)
- DNA ladder (GeneRuler 100bp Plus DNA ladder) used to determine band size
- PCR purification kit (QIAquick PCR purification kit)
- Volume of purified PCR products used for Sanger sequencing (2 μL)
- None specified
- None specified
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