In our study, serial dilutions (1∶2) of the E. coli DNA standard resuspended in TE buffer was extracted with the two extraction methods tested; recovery with MagNA pure compact was done using Total NA plasma 100 400 protocol that produces a final elution volume of 100 µl.
With help of ASTERGDEM of 30 m resolution (source- USGS website) the drainage buffer, slope angle and the aspect maps have been produced whereas the geology, soil and fault buffer maps were prepared with the help of previous published maps (Mahajan and Virdi, 2000) and the land cover and road buffer maps were extracted with help of Google earth imagery.
In brief, 25 mg tissues was homogenised in potassium phosphate buffer, and lipids were extracted with chloroform:methanol (1 : 2).
For protein isolation, actin-motility assays were scaled up (1 ml) then incubated (20 min, RT) before phospholipid-coated beads were isolated by low-speed centrifugation (1000 g), washed ten times in HKSM (20 mM HEPES [pH7.4], 100 mM KCl, 5 mM MgCl2) buffer, and then proteins were extracted with SDS-UREA sample buffer.
The dried seeds were ground to fine powder and the proteins were extracted with extraction buffer [ 12] consisting of 10 mM Na2 HPO4, 15 mM NaH2PO4, 10 mM KCl, and 2 mM EDTA (pH 7.0) by constant stirring overnight at 4°C.
Total protein of mature pollen grains were extracted with extraction buffer (50 mM PBS PH = 7.4, 200 mM NaCl, 10 mM MgCl2, Glycerol 10%, add protease inhibitor cocktail tablets 10 mL/one mini tablet, Roche) and separated by SDS PAGE.
The pellets were washed in TKCM buffer and nuclear proteins were extracted with 2M NaCl in TKCM buffer for 1h at 4°C.
At selected time points, the cells were washed once with cold phosphate buffered saline (PBS), and total proteins were extracted with lysis buffer (1% NP-40, PMSF, 2 mM sodium vanadate, 0.1% sodium deoxycolate, and protease inhibitor cocktail (Roche, Penzberg, Germany).
Cells were harvested at 80 90% confluence, and cellular proteins were extracted with lysis buffer [RIPA buffer, phosphatase inhibitor cocktail 2 and 3 (Sigma-Aldrich), and Complete Mini (Roche, Basel, Switzerland)].
For immunoprecipitation, cells were extracted with buffer A (50 mM Tris, ph 7.4, 150 mM NaCl, 5 mM EDTA, 5 mM EGTA, protease inhibitors) containing 2% SDS at 4°C. for 1 hr.
