Use of Filtron Mini-ultrasette tangential flow device and Filtron Microsep centrifugal concentrators in the early stages of purification of DNA polymerases.
Isolation and initial partial purification of several DNA polymerases from Trypanosoma brucei were achieved. Concentration of relatively large volumes necessary in the early stages of the protocol was accomplished by means of the Filtron Mini-ultrasette tangential flow devices. Subsequently, different size classes of polymerases were obtained by means of Filtron Microsep concentrators, which utilize dead-end filtration and are ideal for concentrating small volumes (several mL) to even smaller volumes (microL). The use of the Filtron systems allowed us to determine relative amounts of different polymerase species quickly and efficiently and to overcome routine problems with concentrating labile macromolecules.
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D000818:Animals; D002498:Centrifugation; D004259:DNA-Directed DNA Polymerase / Q000302:isolation & purification*; D005374:Filtration; D008297:Male; D051381:Rats; D017207:Rats, Sprague-Dawley; D014346:Trypanosoma brucei brucei / Q000201:enzymology*
D004259:DNA-Directed DNA Polymerase