Fluorometric determination of DNA in mammalian cell cultures by PicoGreen

被引:6
作者
Charlton, HR
Relton, JM
Slater, NKH
机构
[1] Aston Univ, Dept Chem Engn & Appl Chem, Birmingham B4 7ET, W Midlands, England
[2] Glaxo Wellcome Res & Dev Ltd, Biotechnol Prod Dev, Purificat Dev, Beckenham BR3 3BS, Kent, England
关键词
DNA quantification; fluorochrome; mammalian cell culture; PicoGreen;
D O I
10.1023/A:1008911713230
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
An intercalating fluorochrome, PicoGreen, was assessed for its ability to determine the concentration of DNA in clarified mammalian cell culture broths containing monoclonal antibodies. Fluorescent signal suppression was ameliorated by sample dilution or by performing the assay above the pI of secreted IgG. The source of fluorescence in clarified culture broth was validated by incubation with RNase A and DNase I. At least 91.8% of fluorescence was attributable to nucleic acid and pre-digestion with RNase A was shown to be a requirement for successful quantification of DNA in such samples.
引用
收藏
页码:681 / 688
页数:8
相关论文
共 16 条
[1]   PicoGreen quantitation of DNA: Effective evaluation of samples pre- or post-PCR [J].
Ahn, SJ ;
Costa, J ;
Emanuel, JR .
NUCLEIC ACIDS RESEARCH, 1996, 24 (13) :2623-2625
[2]   HIGH-LEVEL EXPRESSION OF A RECOMBINANT ANTIBODY FROM MYELOMA CELLS USING A GLUTAMINE-SYNTHETASE GENE AS AN AMPLIFIABLE SELECTABLE MARKER [J].
BEBBINGTON, CR ;
RENNER, G ;
THOMSON, S ;
KING, D ;
ABRAMS, D ;
YARRANTON, GT .
BIO-TECHNOLOGY, 1992, 10 (02) :169-175
[3]   IMPROVED MICRO-FLUOROMETRIC DNA DETERMINATION IN BIOLOGICAL-MATERIAL USING 33258-HOECHST [J].
CESARONE, CF ;
BOLOGNESI, C ;
SANTI, L .
ANALYTICAL BIOCHEMISTRY, 1979, 100 (01) :188-197
[4]  
COOK RJ, 1996, AUTOMATED MICROBIAL, P309
[5]  
Enger O, 1996, BIOTECHNIQUES, V21, P372
[6]   WHO Expert Committee on Biological Standardization - Highlights of the Meeting of October 1996 [J].
Griffiths, E .
BIOLOGICALS, 1997, 25 (03) :359-362
[7]   PICOGRAM QUANTITATION OF TOTAL DNA USING DNA-BINDING PROTEINS IN A SILICON SENSOR-BASED SYSTEM [J].
KUNG, VT ;
PANFILI, PR ;
SHELDON, EL ;
KING, RS ;
NAGAINIS, PA ;
GOMEZ, B ;
ROSS, DA ;
BRIGGS, J ;
ZUK, RF .
ANALYTICAL BIOCHEMISTRY, 1990, 187 (02) :220-227
[8]   DNA, DRAGONS AND SANITY [J].
PETRICCIANI, JC ;
HORAUD, FN .
BIOLOGICALS, 1995, 23 (03) :233-238
[9]   PERIODICITY OF DEOXYRIBONUCLEASE-I DIGESTION OF CHROMATIN [J].
PRUNELL, A ;
KORNBERG, RD ;
LUTTER, L ;
KLUG, A ;
LEVITT, M ;
CRICK, FHC .
SCIENCE, 1979, 204 (4395) :855-858
[10]  
Sambrook J., 2002, MOL CLONING LAB MANU