molecular - nucleic acid extraction and quantification

acid citrate dextrose (yellow top)

what is the preferred tube for molecular biology that has minimal effect on PCR and keeps WBCs alive longer?

sodium heparin (green top)

what is the required tube for cytogenetics?

fresh/snap-frozen tissues

preferred specimen for tissue

neutral buffered formalin or acetone

preferred fixative for fixed tissues

extracted DNA

storage 4C for short term and -20C for long term

extracted RNA

storage -80C and under LN2 is better

microdissection

allows for concentration of specific tissues or areas of tissue in extraction, scalpel dissection, microscope assisted or laser assisted

phenol chloroform

what is the organic reagent for liquid extraction for DNA?

acid guaninidium isothiocyonate-phenol-chloroform

what is the organic reagent for liquid extraction for RNA?

liquid extractions (organic)

aqueous lysate mixed with organic reagent, nucleic acids in top aqueous layer and proteins at interface

salting out (inorganic)

proteins precipitated with high salt solutions

solid phase extraction

uses silica membrane or silica coated magnetic beads, DNA binds to that in presence of high ionic strength solution, eluted with low salt solution

260nm

nucleic acids absorb best at _____

50 ug/mL

dsDNA concentration factor

40 ug/mL

RNA concentration factor

dye binding assay

DNA quantification where fluorescent dye is added to sample and signal is measured

270-275nm

phenol contamination range

280nm

protein contamination range

230nm

salts and organic solvents range

gel electrophoresis

can be used to assess whether DNA is mostly intact or degraded