Week 7 - Nucleic Acid Extraction

Miescher

first isolated DNA in human cells in 1869 by precipitation

Density-gradient centrifugation strategies

DNA isolation techniques were developed from?

Meselson and Stahl

1958, demonstrate semiconservative replication of DNA

Enzymes

Glass beads

Detergent

Strong base (0.2M NaOH in the presence of triEDTA)

Glucose

Bacteria and fungi have tough cell walls that can be broken by?

Boiling in dilute sucrose, triton X-100 detergent, tris buffer, and EDTA after lysozyme treatment releases DNA that can be immediately precipitated with alcohol.

Boiling in _________, __________, __________, and EDTA after lysozyme treatment releases DNA that can be immediately precipitated with alcohol.

Denatured

DNA extracted by boiling or subjected in alkaline is ____ and may not be suitable for restriction enzyme analysis that require double-stranded DNA.

Held within free viruses or integrated into the host genome along with host DNA

How are viruses prepared for analysis?

Differential density gradient centrifugation or Differential lysis

WBCs are purified from other blood components by?

Differential density-gradient centrifugation

Whole blood or bone marrow sample mixed with isotonic saline overlaid with Ficoll

Ficoll

Highly branched sucrose polymer that does not penetrate biological membranes

Plasma

Solid tumors and transplanted organs release cells, exosomes, and nucleic acid into the bloodstream