Protocol
1) Take 1.5ml of the bacterial culture, centrifuge at 14,000 rpm for 5mins.
2) Decant the supernatant completely & make a single cell suspension.
3) Add 500ul of 10mM Tris, 10mM EDTA, 100mM NaCl, 2% SDS, 20ug/ml of proteniase K.
4) Mix well and incubate at 560C for 30mins.
5) Add 500ul isopropanol, mix well & centrifuge at 14,000rmp for 10mins.
6) Decant the supernatant & add 200ul of 80% ethanol, centrifuge at 14,000rpm for 10mins at 40C.
7) Repeat the above step.
8) Air dry the pellet.
9) Dissolve in 200ul of sterile distil water.
10) Run the samples on 0.8% agarose gel.
Note: Phenol Chloroform purification can be performed to get pure DNA preparation.
1) Take 1.5ml of the bacterial culture, centrifuge at 14,000 rpm for 5mins.
2) Decant the supernatant completely & make a single cell suspension.
3) Add 500ul of 10mM Tris, 10mM EDTA, 100mM NaCl, 2% SDS, 20ug/ml of proteniase K.
4) Mix well and incubate at 560C for 30mins.
5) Add 500ul isopropanol, mix well & centrifuge at 14,000rmp for 10mins.
6) Decant the supernatant & add 200ul of 80% ethanol, centrifuge at 14,000rpm for 10mins at 40C.
7) Repeat the above step.
8) Air dry the pellet.
9) Dissolve in 200ul of sterile distil water.
10) Run the samples on 0.8% agarose gel.
Note: Phenol Chloroform purification can be performed to get pure DNA preparation.
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