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Table 2 DNA extraction protocols for the four sample preservation methods

From: Low-cost sample preservation methods for high-throughput processing of rumen microbiomes

Method

Sample defrosting time

Sample amount used

Rumen sample used (DM) (mg)

Solution (µL)

Amount of supernatant taken5 (µL)

Buffer A1

Buffer PM2

20% SDS3

P:C:I4

GRC

N/A

30 mg

30

282

268

200

550

 ~ 350

TNx2

15 min

1 mL

34.5

N/A

268

N/A

550

500

GHx2

10 min

1 mL

34.5

N/A

268

N/A

550

500

EtOH

5 min

0.8 mL

18.2

N/A

268

200

550

 < 350

  1. N/A denotes not applicable, and DM denotes dry matter
  2. 1200 mM NaCl, 200 mM Tris, 20 mM EDTA, pH 8 adjusted with NaOH
  3. 2Binding buffer (Qiagen)
  4. 3Sodium dodecyl sulfate (20% wt/vol)
  5. 4Phenol/chloroform/isoamyl alcohol
  6. 5Supernatant taken after 4 min of bead beating and centrifugation at 16,060 g and 4 °C for 20 min
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Animal Microbiome

ISSN: 2524-4671