Chromatography in DNA radiolabeling: hands-off automation using a robotic workstation.
The experiments described in the present paper were performed in order to determine whether the Biomek-1000 (Beckman Instruments, Fullerton, CA, USA) automated laboratory workstation can be used in a fully automated DNA labeling method followed by automated gravity-driven size exclusion purification of molecular probes. To this aim, we performed random oligodeoxyribonucleotide priming of a HIV-1 LTR probe that was used for molecular hybridization to Southern blotted polymerase chain reaction products. The results obtained demonstrate that the automatically labeled probe can be efficiently purified by automated and gravity-driven Sephadex G-50 chromatography, without any major changes in hybridization property. This robotic methodology can be used in several procedures employing radioisotope labeling.
Mischiati
C
C
Department of Biochemistry and Molecular Biology, Ferrara University, Italy.
Feriotto
G
G
Fiorentino
D
D
Gambari
R
R
D001483:Base Sequence; D015139:Blotting, Southern; D002460:Cell Line; D002850:Chromatography, Gel; D004247:DNA / Q000737:chemistry*; D015342:DNA Probes; D015497:HIV-1 / Q000235:genetics; D006684:HLA-DR Antigens / Q000235:genetics; D006801:Humans; D017403:In Situ Hybridization; D007553:Isotope Labeling / Q000295:instrumentation*; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D012371:Robotics; D013601:T-Lymphocytes / Q000378:metabolism
D015342:DNA Probes; D006684:HLA-DR Antigens; D004247:DNA