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Table 3 Results of validation study of species-specific heminested PCR assays

From: Development of novel heminested PCR assays based on mitochondrial 16s rRNA gene for identification of seven pecora species

Validation Parameters No. of samples analysed No. of specific primers analysed No. of samples successfully typed
DNA concentration 5pg 20 7 20
  50 pg 20 7 20
  500 pg 20 7 20
  5 ng 20 7 20
Mixture analysis $ Blackbuck + Nilgai + Goral + Hog deer + Chital + Thamin + Sambar 1 (mixture) 1a 1 (Blackbuck)
  Blackbuck + Nilgai + Goral + Hog deer + Chital + Thamin + Sambar 1 (mixture) 1b 1 (Nilgai)
  Blackbuck + Nilgai + Goral + Hog deer + Chital + Thamin + Sambar 1 (mixture) 1c 1 (Goral)
  Blackbuck + Nilgai + Goral + Hog deer + Chital + Thamin + Sambar 1 (mixture) 1d 1 (Hog deer)
  Blackbuck + Nilgai + Goral + Hog deer + Chital + Thamin + Sambar 1 (mixture) 1e 1 (Chital)
  Blackbuck + Nilgai + Goral + Hog deer + Chital + Thamin + Sambar 1 (mixture) 1f 1 (Thamin)
  Blackbuck + Nilgai + Goral + Hog deer + Chital + Thamin + Sambar 1 (mixture) 1g 1 (Sambar)
Chemical reagents Soap 20 7 20
  0.1 N NaOH 20 7 20
  5% Acetic acid 20 7 20
  1 M NaCL 20 7 20
  Gasoline 20 7 20
Exposure to Heat 37°C, 1 week 7* 7 7
  37°C, 2 week 7* 7 7
  37°C, 3 week 7* 7 7
UV Irradiation (302 nm, 15 cm distance) 1 h 20 7 20
  8 h 20 7 20
  24 h 20 7 18
  1. * One sample of each species were included for Exposure to Heat validation
  2. $ Each mixture comprised 50 pg of DNA of target species and 500 pg of DNA of each non-target species a – Blackbuck specific primer, b – Nilgai specific primer, c- Goral specific primer, d – Hog deer specific primer, e – Chital specific primer, f – Thamin specific primer, g – Sambar specific primer.