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 ðanol
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