Isolation of Genomic DNA from Scarlet Runner Bean Leaves Using DNeasy Plant Mini

Kit

HC 70ALSPRING 2009APRIL 7, 2009

ANHTHU BUI

Isolation of Genomic DNA from Scarlet Runner Bean Leaves Using DNeasy Plant Mini

KitSRB Leaves

Freezed in liquid nitrogenGrind into powder

with a mortar & pestle

Frozen SRB Leaf Powder

Lyse cells with AP1 extraction buffer

Genomic DNA, ribosomal RNAs, tRNAs, mRNAs, proteins,

polysaccharidesDigest RNAs with RNase A

Centrifuge

Genomic DNA, proteins, oligo-ribonucleotides, polysaccharides

Filtered & homogenizedvia Shredder column

Adjust buffer conditions byadding AP3 buffer &ethanol

Lysate

Lysate

Cell debris

Separate DNA from contaminants using a silica-gel spin column

Genomic DNA bound to silica-gel column

Wash column

Centrifuge

Elute with water or low-salt buffer

Pure genomic DNA

Precipitate polysaccharides & proteins with AP2 buffer