Bioline ISOLATE II Plant DNA Kit, Product Manual

Page: 6 / 20

Product Manual www.bioline.com/isolate
6
Plant DNA Isolation
Plant sample lysis
Add 400 µL Lysis Buffer
PA1
10 µL RNase A Mix
Incubate 65
o
C, 10 min
11,000 x g, 1 min
Homogenize plant sample
100 mg tissue, 20 mg dried
Plant sample lysis
Add 300 µL Lysis Buffer
PA2
10 µL RNase A Mix
Incubate 65
o
C, 10 min
Add 75 µL Precipitation Buffer
PL3
Incubate on ice, 5 min
Filter crude lysate
11,000 x g, 2 min
1 st and 2 nd
11,000 x g, 1 min
Adjust DNA binding conditions
Add 450 µL Binding Buffer PB
Vortex
Bind DNA
Load lysate
3 rd
11,000 x g, 2 min
Wash and dry silica membrane
1
st
wash 400 µL Wash Buffer PAW1
2
nd
wash 700 µL Wash Buffer PAW2
3 rd wash 200 µL Wash Buffer PAW2
11,000 x g, 1 min
Repeat DNA elution step
Isolated DNA
Elute DNA
Add 50 µL Elution Buffer PG (65
o
C)
Incubate 65
o
C, 5 min