lab.rogierstuger.com
MOLECULAR CELL ALCHEMY COOKBOOK


solutions

antibiotics stock solutions | general stock solutions


culture media

culture media: yeast and E. coli

culture media: plant cells
(arabidopsis thaliana suspension culture, tomato suspension culture, tobacco leaf protoplast or cultures made thereof)


general methods

DNA size markers from plasmids | RNA size markers by in vitro transcription

glycerol stocks of E. coli


DNA methods + cloning

transformation

E. coli (fast and very very simple) | E. coli (SEM method, simple and efficient)
tobacco leaf protoplasts

yeast (Saccharomyces cerevisiae)

plasmid DNA preps

qiagen miniprep and column recycle protocol tip20 | tip100
quick'n'dirty miniprep

yeast miniprep

DNA precipitation & cleanup

precipitation: ethanol | isopropanol

DNA cut'n'paste methods

restriction enzymes | ligation

PCR

PCR on E. coli clones | PCR on yeast clones

DNA electrophoresis

agarose gel electrophoresis
chloroquin-agarose electrophoresis for resolution of supercoiled topoisomers

Southern blotting

transfer, probing and detection with DIG-labeled probes


RNA methods

RNA isolation from plant cells
RNA synthesis by in vitro transcription

RNA precipitation & cleanup

RNA electrophoresis

acrylamide/urea minigel format (fast) | sequencing gel format (high resolution of small RNA molecules)


protein methods + western blots

protein precipitation with TCA, ammonium sulphate, acetone, ethanol

protein electrophoresis

SDS-PAGE minigels
Schagger-von Jagow (high resolution of very small peptides)

protein quantitation

whole cell protein assay (modified Lowry method)
spot blot quantitation of proteins in blue SDS-PAGE buffer (using amido black)

Western blotting

staining total protein on blots with Ponceau S, amido black, coomassie blue
detection by primary and secondary antibodies

stripping Western blots


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