Table 2 Comparison of rHTTP method with other reported methods of quick plant DNA extraction
Rapid One Step Extraction (ROSE) method (Steiner et al., 1995) | Sucrose Prep Method (Berendzen et al. 2005) | NaOH Method (Wang et al., 1993) | NaOH Method (Satya et al. 2013) | NaOH Method (Wang et al., 2015) | rHTTP method (Present study) | |
|---|---|---|---|---|---|---|
Type and form of plant material | Lyophilized plant tissue | Fresh tissue | Young tissue | Fresh tissue | Fresh tissue | Fresh/old tissue |
Liquid nitrogen | No | Yes/Ice | No | No | No | No |
Extraction/lysis buffer | Tris-HCl, EDTA, Sodium Sarkosyl sulfate, PVPP | Tris-HCl, NaCl, Sucrose | NaOH, Tris | NaOH, Tris-EDTA | NaOH | SDS |
Instrumentation | Lyophilizer, Shaker, hybridization Oven/ Water bath | Water Bath/Thermal cycler, Centifuge, grinding assembly/Mortar pestle | Grinding assembly/ Mortar-pestle | Grinding assembly/ Mortar-pestle, Centrifuge | Grinding assembly/ Mortar-pestle/ centrifuge | Thermal cycler/hot plate/any means to boil water |
Duration to make templates for PCR (as per original publication) | ~ 45 min after lyophilization | ~ 15 min | Should not be more than 10 min | 10–12 min | 5 min | 10 min |
Actual time required to process five samples at a time | ND | 59 min (11.8 min per sample) *For larger number of samples time will increase proportionately due to grinding | 31 min (6.2 min per sample) *For larger number of samples time will increase proportionately due to grinding | 65 min (13 min per sample) *For larger number of samples time will increase proportionately due to grinding | 29 min (5.8 min per sample) *For larger number of samples time will increase proportionately due to grinding | 12.5 min (2.5 min per sample) *For larger number of samples, the time will remain almost same as there is no grinding or maceration. The time will be 10 min plus time required for pipetting 100 μl lysis buffer |