Set up 20 µl PCR reactions with template DNA (82.5 ng )10 mM Tris-HCl, , . #####%%%%% output "primers" are used in line6
{"action": "set", "output": "primers", "reagent": ["template DNA (82.5 ng)", "10 mM Tris-HCl"], "volume": ["20 µl"]}
Run the PCR program: 94°C (2 min), 94°C (1 min), 60°C (1 min), 72°C (1.75 min) for 35 cycles, followed by 72°C (10 min).
{"action": "run", "output": "", "reagent": [""], "temperature": ["94°C", "94°C", "60°C", "72°C"], "time": ["2 min", "1 min", "1 min", "1.75 min"]}
Detect DH-JH recombination as amplified fragments (1,033 bp, 716 bp, 333 bp) using primers DHL(5’) . #####%%%%% output "germline alleles" are used in line8
{"action": "detect", "output": "germline alleles", "reagent": ["DH-JH recombination", "primers", "DHL5\u2019"]}
Detect germline alleles as a 1,259 bp fragment using primers Mu0(5’) .
{"action": "detect", "output": "", "reagent": ["germline alleles", "primers"]}