16S rDNA sequences of soil samples in alpine meadow and alpine steppe meadow
Soil microbial DNA extraction was obtained and purified. V4 hypervariable region of bacterial 16S rDNA was amplified. The reverse primer combined with a barcode sequence. PCR amplification was used in a 25 ¦Ìl reaction, containing 1 ¦Ìl of each primer, 2.5 ¦Ìl AccuPrime PCR buffer II (Invitrogen, Grand Island, NY, USA), 5 ¦Ìl DNA and 0.1 ¦Ìl AccPrime Taq Polymerase. The reaction mixture was denatured at 94 oC for 1 min, followed by 30 cycles of 94 oC for 20 s, 53 oC for 25 s and 68 oC for 45s, and extension at 68 oC for 10 min. The PCR products were purified and run using a Miseq (Illumina, San Diego, CA, USA).
SUB2385159
SJYAM8