Effect of temperature and 4,6'-diamidine-2-phenylindole on restriction of supercoiled Col E1 DNA by Eco RI endonuclease.
Supercoiled Col E1 DNA is split by Eco RI endonuclease at 37 degrees C without intermediate formation of open circular DNA. Accumulation of this restriction product is observed at low temperature. The fluorescent dye, 4,6'-diamidine-2-phenylindole (DAPI) inhibits restriction by Eco RI endonuclease. This effect is due to the DAPI:DNA rather than to the DAPI:Eco RI interactions.
D000578:Amidines / Q000494:pharmacology; D004262:DNA Restriction Enzymes / Q000037:antagonists & inhibitors / Q000235:genetics*; D004269:DNA, Bacterial / Q000378:metabolism*; D004270:DNA, Circular / Q000096:biosynthesis; D004278:DNA, Superhelical / Q000378:metabolism*; D003864:Depression, Chemical; D004926:Escherichia coli / Q000235:genetics*; D005456:Fluorescent Dyes / Q000494:pharmacology*; D007211:Indoles / Q000494:pharmacology*; D010957:Plasmids; D013696:Temperature
D000578:Amidines; D004269:DNA, Bacterial; D004270:DNA, Circular; D004278:DNA, Superhelical; D005456:Fluorescent Dyes; D007211:Indoles; C007293:DAPI; D004262:DNA Restriction Enzymes