DNA sequencing with dye-labeled terminators and T7 DNA polymerase: effect of dyes and dNTPs on incorporation of dye-terminators and probability analysis of termination fragments.
The incorporation of fluorescently labeled dideoxynucleotides by T7 DNA polymerase is optimized by the use of Mn2+, fluorescein analogs and four 2'-deoxyribonucleoside 5'-O-(1-thiotriphosphates) (dNTP alpha S's). The one-tube extension protocol was tested on single-stranded templates, as well as PCR fragments which were made single-stranded by digestion with T7 gene 6 ... More
Injection of ultrasmall samples and single molecules into tapered capillaries.
AuthorsChiu DT, Hsiao A, Gaggar A, Garza-López RA, Orwar O, Zare RN
JournalAnal Chem
PubMed ID9164157
A novel injection scheme is described in which ultrasmall samples in the attoliter (10(-18) L) and low femtoliter (10(-15) L) range, or even single molecules, are controllably introduced into a tapered capillary so that electrophoretic separation can be carried out. To match the dimensions of the capillary inlet with that ... More
Nanoengineered structures for holding and manipulating liposomes and cells.
AuthorsWilson CF, Simpson GJ, Chiu DT, Strömberg A, Orwar O, Rodriguez N, Zare RN
JournalAnal Chem
PubMed ID11248893
We describe the fabrication of nanoengineered holding pipets with concave seating surfaces and fine pressure control. These pipets were shown to exhibit exceptional stability in capturing, transporting, and releasing single cells and liposomes 1-12 microm in diameter, which opens previously inaccessible avenues of research. Three specific examples demonstrated the utility ... More
Applications of fluorescence correlation spectroscopy: measurement of size-mass relationship of native and denatured schizophyllan.
AuthorsLeng X, Starchev K, Buffle J
JournalBiopolymers
PubMed ID11473353
Diffusion dynamics of a polysaccharide, schizophyllan has been studied by fluorescence correlation spectroscopy (FCS). Several different sizes of nondenatured and denatured schizophyllan have been labeled with rhodamine 6G in borate buffer. The length of the nondenatured schizophyllan was calculated from FCS data by using the Broersma's relationship for rod-like macromolecules. ... More
Four-color DNA sequencing by synthesis on a chip using photocleavable fluorescent nucleotides.
AuthorsSeo TS, Bai X, Kim DH, Meng Q, Shi S, Ruparel H, Li Z, Turro NJ, Ju J
JournalProc Natl Acad Sci U S A
PubMed ID15829588
We report four-color DNA sequencing by synthesis (SBS) on a chip, using four photocleavable fluorescent nucleotide analogues (dGTP-PC-Bodipy-FL-510, dUTP-PC-R6G, dATP-PC-ROX, and dCTP-PC-Bodipy-650) (PC, photocleavable; Bodipy, 4,4-difluoro-4-bora-3alpha,4alpha-diaza-s-indacene; ROX, 6-carboxy-X-rhodamine; R6G, 6-carboxyrhodamine-6G). Each nucleotide analogue consists of a different fluorophore attached to the 5 position of the pyrimidines and the 7 position ... More