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 |