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Table 1 Comparison of DNA extraction methods

From: Extraction of high purity genomic DNA from pine for use in a high-throughput Genotyping Platform

Method

Wt. of tissue (mg)

Homo-genisation conditions

Extraction buffer used(vol; temp)

Lysis incubation conditions

Centrifug-ation conditions

RNase A conc.

Incubation 2 conditions

Extraction conditions

DNA pptn conditions

Additional purification

S

300

Pestle & mortar under liquid nitrogen

1 mL CTAB buffer; 65°C

60 min; 65°C

18000× g; 20 min

100 μg/ mL

30 min; 37°C (RNase A digestion)

(1) 0.2 vol 5 M NaCl + 1 vol CIA1; then 18000× g 20 min

1 vol cold isopropanol; -20°C O/N3; then 18000× g 30 min

None

(2) 1 vol CIA1; then 18000× g 20 min

C

150

Geno/ GrinderTM 2000 with sea sand, 2 stainless steel beads

2 mL CTAB buffer; RT2

Same as for Method S

2000× g; 10 min

100 μg/ mL

Same as for Method S

Same as for Method S

1 vol RT2 isopropanol; RT2 O/N3; then 18000× g 30 min

None

CE

Same as for Method C

       

Ethanol/sodium acetate

CQ

Same as for Method C

       

QIAquick® PCR Purification kit

CZ

Same as for Method C

       

Genomic DNA Clean and Concentrator™ kit

CG

Same as for Method C

       

Genomic-tip 20/G kit

F

Same as for Method C

FastPrep® instrument, tubes with beads supplied in kit

Kit supplied

5 min; on ice

Same as for Method S

None

N/A

Kit supplied

N/A

None

D & D+

50

Same as for Method C

Kit supplied

N/A

Same as for Method C

250 μg/ mL

10 min; -20°C (SDS precipitation)

Kit supplied

N/A

None

N & N96

100

Same as for Method C

Kit supplied

Same as for Method S

Same as for Method C

300 μg/ mL

Same as for Method D

Kit supplied

N/A

None

  1. 1 Chloroform:isoamyl alcohol (24:1) vol:vol.
  2. 2 Room temperature.
  3. 3 Overnight.