Fluorescently labeled model DNA sequences for exonucleolytic sequencing.
AuthorsFöldes-Papp Z, Angerer B, Thyberg P, Hinz M, Wennmalm S, Ankenbauer W, Seliger H, Holmgren A, Rigler R
JournalJ Biotechnol
PubMed ID11257532
'We describe here the enzyme-catalyzed, low-density labeling of DNAs with fluorescent dyes. Firstly, for "natural" template DNAs, dNTPs were partially substituted in the labeling reactions by the respective fluorophore-bearing analogs. The DNAs were labeled by PCR using Taq DNA polymerase. The covalent incorporation of dye-dNTPs decreased in the following order: ... More
In vitro production and screening of DNA polymerase eta mutants for catalytic diversity.
AuthorsGlick E, Anderson JP, Loeb LA
JournalBiotechniques
PubMed ID12449395
'Mutant DNA polymerases have become an increasingly important tool in biotechnology. The ability to examine the activity and specific properties of enzymes has a crucial role in the characterization of the enzyme. We have developed several systems for characterizing DNA polymerases that combine random mutagenesis with in vivo selection systems. ... More
RNA structure analysis assisted by capillary electrophoresis.
AuthorsSobczak K, Krzyzosiak WJ
JournalNucleic Acids Res
PubMed ID12434006
'Although most capillary electrophoresis (CE) applications in the nucleic acid field performed so far address DNA analysis, various RNA assays by CE have also been done. Both natural and synthetic RNAs have been examined to evaluate their quantities, sizes and interactions. In this report, we show a novel application of ... More
High-density polymerase-mediated incorporation of fluorochrome-labeled nucleotides.
AuthorsRamanathan A, Pape L, Schwartz DC
JournalAnal Biochem
PubMed ID15649370
'DNA-polymerase-mediated incorporation of different fluorochrome-labeled nucleotides (FdNTPs) was investigated with the goals of optimizing the high-density labeling of probes and exploring DNA sequencing strategies that rely on the controlled, sequential addition of such compounds. By systematically evaluating variables--including polymerase type, buffer conditions, and fluorochrome chemistries--a rational strategy for the sequential ... More
Real-time enzyme kinetics monitored by dual-color fluorescence cross-correlation spectroscopy.
AuthorsKettling U, Koltermann A, Schwille P, Eigen M
JournalProc Natl Acad Sci U S A
PubMed ID9465029
'A method for sensitively monitoring enzyme kinetics and activities by using dual-color fluorescence cross-correlation spectroscopy is described. This universal method enables the development of highly sensitive and precise assays for real-time kinetic analyses of any catalyzed cleavage or addition reaction, where a chemical linkage is formed or cleaved through an ... More
Telomere length dynamics and chromosomal instability in cells derived from telomerase null mice.
AuthorsHande MP, Samper E, Lansdorp P, Blasco MA
JournalJ Cell Biol
PubMed ID10037783
To study the effect of continued telomere shortening on chromosome stability, we have analyzed the telomere length of two individual chromosomes (chromosomes 2 and 11) in fibroblasts derived from wild-type mice and from mice lacking the mouse telomerase RNA (mTER) gene using quantitative fluorescence in situ hybridization. Telomere length at ... More
Spectral karyotyping combined with locus-specific FISH simultaneously defines genes and chromosomes involved in chromosomal translocations.
AuthorsTonon G, Roschke A, Stover K, Shou Y, Kuehl WM, Kirsch IR
JournalGenes Chromosomes Cancer
PubMed ID10719373
Genes that play roles in malignant transformation have often been found proximate to cancer-associated chromosomal breakpoints. Identifying genes that flank chromosomal reconfigurations is thus essential for cancer cytogenetics. To simplify and expedite this identification, we have developed a novel approach, based on simultaneous spectral karyotyping and fluorescence in situ hybridization ... More