Localization of trinucleotide repeat sequences in myotonic dystrophy cells using a single fluorochrome-labeled PNA probe.

abstract：:Here we report the construction of a histidine-tagged T4 RNA ligase expression plasmid (pRHT4). The construct, when overexpressed in BL21 (DE3) cells, allows the preparation of large quantities of T4 RNA ligase in high purity using only a single purification column. The histidine affinity tag does not inhibit enzyme f...

journal_title：BioTechniques

authors： Wang QS,Unrau PJ

更新日期：2002-12-01 00:00:00

abstract：:This report describes a new method for labeling PCR-generated short length (60-120 bp) double-stranded DNA fragments for use as hybridization probes. The method utilizes gene-specific primers identical to those for PCR generation of non-radioactive DNA fragments. Radioactive probes are synthesized by Taq DNA polymeras...

journal_title：BioTechniques

authors： Mizobuchi M,Frohman LA

更新日期：1992-03-01 00:00:00

abstract：:The limiting detection signal for identification of human genetic markers, such as HLA-D and VNTR genes, was determined using DNA isolated from a series of decreasing numbers of lymphocytes carrying the target marker in the polymerase chain reaction (PCR). The PCR procedure was assembled by incorporating 32P-labeled d...

journal_title：BioTechniques

authors： McDaniel DO,Naftilan J,Barber WH

更新日期：1993-07-01 00:00:00

abstract：:Piezoelectric dispensing of proteins from borosilicate glass capillaries is a popular method of protein biochip fabrication that offers the advantages of sample recovery and noncontact with the printing substrate. However, little regard has been given to the quantitative aspects of dispensing minute volumes (1 nL or l...

journal_title：BioTechniques

authors： Delehanty JB,Ligler FS

更新日期：2003-02-01 00:00:00

abstract：:Cell culture is a vital component of laboratories throughout the scientific community, yet the absence of standardized protocols and documentation practice challenges laboratory efficiency and scientific reproducibility. We examined the effectiveness of a cloud-based software application, CultureTrax® as a tool for st...

journal_title：BioTechniques

authors： Canfield SG,Jin G,Palecek SP,Sampsell T

更新日期：2018-11-01 00:00:00

abstract：:We describe a rapid and sensitive method for the detection of nucleotide sequence variation that can be used for large-scale screening of population markers. Denaturing gradient gel electrophoresis (DGGE) detects sequence variants of amplified fragments by the differences in their melting behavior. DGGE detects most s...

journal_title：BioTechniques

authors： Miller KM,Ming TJ,Schulze AD,Withler RE

更新日期：1999-11-01 00:00:00

abstract：:The increase of information in biology makes it difficult for researchers in any field to keep current with the literature. The MEDLINE database of scientific abstracts can be quickly scanned using electronic mechanisms. Potentially interesting abstracts can be selected by matching words joined by Boolean operators. H...

journal_title：BioTechniques

authors： Perez-Iratxeta C,Keer HS,Bork P,Andrade MA

更新日期：2002-06-01 00:00:00

abstract：:A new method has been developed to purify and detritylate milligram amounts of synthetic oligonucleotides. Dimethoxytrityl oligonucleotides from 15 to 100 nucleotides in length are applied in triethylammonium acetate or concentrated ammonium hydroxide to a disposable chromatographic cartridge, the NENSORB PREP Nucleic...

journal_title：BioTechniques

authors： Johnson BA,McClain SG,Doran ER,Tice G,Kirsch MA

更新日期：1990-04-01 00:00:00

abstract：:The two-hybrid system in Saccharomyces cerevisiae is a genetic approach for the detection of of protein-protein interactions in vivo. This technology relies on the the activity of separated DNA-binding and transactivation domains of specific transcription factors to reconstitute an active transcription factor complex ...

journal_title：BioTechniques

authors： Mayer G,Launhardt H,Munder T

更新日期：1999-07-01 00:00:00

abstract：:The relative spatial distribution of cells in a solid tumor contributes to development of malignancy, yet the details of this process remain poorly understood. To elucidate these mechanisms, the ability to extract and analyze the entire DNA content of individual cells whose precise location in the tumor is known is re...

journal_title：BioTechniques

authors： Guo Y,Yang Y,Zhou J,Czajkowsky DM,Liu B,Shao Z

更新日期：2012-04-01 00:00:00

abstract：:A low order neural network-based filter was designed as a rapid screening agent for single-spanning transmembrane regions in an integrated informatics system. A rapid screening algorithm was seen as a compromise between costly structure-specific techniques and simple rules that gave a high false-positive rate for cDNA...

journal_title：BioTechniques

authors： Huang GM,Farkas J,Hood L

更新日期：1996-12-01 00:00:00

abstract：:Oligonucleotide primers used to amplify target DNA regions via PCR should meet certain design criteria to maximize the potential for efficient priming. The Random Oligonucleotide Construction Kit (ROCK), a spreadsheet-based program that runs under Microsoft Excel 97 or later version for Microsoft Windows, was develope...

journal_title：BioTechniques

authors： Strain SR,Chmielewski JG

更新日期：2001-06-01 00:00:00

abstract：:A technique that can be used to isolate vector/insert junctions from clones in vectors, such as yeast artificial chromosomes and P1s, and to sequence plasmid inserts more rapidly has been developed. A vector primer is combined with single, randomly chosen oligonucleotides in PCRs, to create pools of products. With 12-...

journal_title：BioTechniques

authors： Swensen J

更新日期：1996-03-01 00:00:00

abstract：:Biopharmaceutical products are of great importance in the treatment or prevention of many diseases and represent a growing share of the global pharmaceutical market. The usual technology for protein synthesis (cell-based expression) faces certain obstacles, especially with 'difficult-to-express' proteins. Cell-free pr...

journal_title：BioTechniques

authors： Chiba CH,Knirsch MC,Azzoni AR,Moreira AR,Stephano MA

更新日期：2021-01-20 00:00:00

abstract：:The ZEBRA protein is a transcriptional activator that induces expression of viral lytic genes in cells harboring latent Epstein-Barr virus (EBV). In this report it is shown that a derivative of ZEBRA that cannot activate transcription (Zd) can be used to detect and characterize activation domains. Three expression vec...

journal_title：BioTechniques

authors： Asković S,Baumann R

更新日期：1997-05-01 00:00:00

abstract：:High-throughput approaches for gene cloning and expression require the development of new nonstandard tools for molecular biologists and biochemists. We introduce a Web-based tool to design primers specifically for the generation of expression clones for both laboratory-scale and high-throughput projects. The applicat...

journal_title：BioTechniques

authors： Yoon JR,Laible PD,Gu M,Scott HN,Collart FR

更新日期：2002-12-01 00:00:00

abstract：:We have developed a new strategy for differential screening of genes that are expressed in two or more tissues, and have used it to identify genes that are preferentially expressed in both testis and ovary. In this approach, testis-specific cDNAs were first generated using the suppression subtractive hybridization tec...

journal_title：BioTechniques

authors： Jin H,Cheng X,Diatchenko L,Siebert PD,Huang CC

更新日期：1997-12-01 00:00:00

abstract：:Cell-based immunizations are often used when membrane antigens are difficult to purify. To confirm that an antibody binding to the surface of a cell line is, in fact, binding to the desired antigen, FACS can be performed independently on two cell lines, a transfected cell line expressing the antigen of interest and a ...

journal_title：BioTechniques

authors： Yang XP,Gallo M,Ngan I,Nocerini M,Chen MM

更新日期：2002-03-01 00:00:00

abstract：:An improved method was developed for in situ reverse transcription-polymerase chain reaction (RT-PCR) to detect and localize mRNA in tissue sections. The coverslip mounted-immersion cycled (COSMIC) in situ RT-PCR technique combines the advantages of solution-phase PCR with the tissue immobilization necessary for in si...

journal_title：BioTechniques

authors： Ertsey R,Scavo LM

更新日期：1998-01-01 00:00:00

abstract：:We have designed and built a magnetic tweezers device that enables the application of calibrated stresses to soft materials while simultaneously measuring their microscale deformation using confocal microscopy. Unlike previous magnetic tweezers designs, our device is entirely portable, allowing easy use on microscopes...

journal_title：BioTechniques

authors： Yang Y,Lin J,Meschewski R,Watson E,Valentine MT

更新日期：2011-07-01 00:00:00

abstract：:Monitoring morphogenetic processes, at high resolution over time, has been a long-standing goal of many developmental cell biologists. It is critical to image cells in their natural environment whenever possible; however, imaging many warm-blooded vertebrates, especially mammals, is problematic. At early stages of dev...

journal_title：BioTechniques

authors： Rupp PA,Rongish BJ,Czirok A,Little CD

更新日期：2003-02-01 00:00:00

abstract：:Bovine serum albumin, free of deoxyribonuclease activity, was obtained in our laboratory using ion-exchange chromatography followed by acetylation. Chromatography on four different resins (DEAE-52, P-11, hydroxylapatite and Q Sepharose fast-flow) was examined. Fractions from Q Sepharose chromatography, eluted with a l...

journal_title：BioTechniques

authors： Trujillo LE,Castellanos L,Garcia O,Herrera L

更新日期：1990-11-01 00:00:00

abstract：:Knob heterochromatin served as the model for the development of fluorescent chromosome in situ hybridization on maize meiotic chromosomes. The meiotic chromosomes were hybridized with a digoxigenin-labeled RNA probe of the knob repeat sequence that is a component of the morphologically determined knob heterochromatin....

journal_title：BioTechniques

authors： Makowski ER,Ruzin SE

更新日期：1994-02-01 00:00:00

abstract：:Intrinsically disordered proteins (IDPs) are subject to post-translational modifications. This allows the same polypeptide to be involved in different interaction networks with different consequences, ranging from regulatory signalling networks to the formation of membrane-less organelles. We report a robust method fo...

journal_title：BioTechniques

authors： Brázda P,Šedo O,Kubíček K,Štefl R

更新日期：2019-07-01 00:00:00

abstract：:A method was developed for the detection of bacterial mRNAs using reverse transcriptase followed by the polymerase chain reaction (PCR) and Southern blot analysis. The method involves brief inhibition of protein synthesis with chloramphenicol, followed by reverse transcription, PCR amplification of cDNA and Southern b...

journal_title：BioTechniques

authors： Mahbubani MH,Bej AK,Miller RD,Atlas RM,DiCesare JL,Haff LA

更新日期：1991-01-01 00:00:00

abstract：:SNPCEQer identifies and reports SNPs in sequences obtained from the Beckman CEQ2000 DNA Analysis System. SNPCEQer aligns sequences obtained using CEQ2000 heterozygote detection analysis and reports discrepancies between individual sequences and the consensus sequence it generates from this set as SNPs when the individ...

journal_title：BioTechniques

authors： Flood EM,Tang F,Horvath MM,Pertsemlidis A,Garner HR

更新日期：2002-10-01 00:00:00

abstract：:Sequencing of the human genome in combination with computational annotation has provided tremendous data on predicted genes. However, for most of them, no corresponding cDNAs are available yet. Furthermore, even where cDNA clones were obtained, gene transcripts often have many different splice variants that are not co...

journal_title：BioTechniques

authors： Mitani Y,Nakayama T,Harbers M,Hayashizaki Y

更新日期：2004-07-01 00:00:00

abstract：:Cysts of free-living protozoa have an impact on the ecology and epidemiology of bacteria because they may act as a transmission vector or shelter the bacteria against hash environmental conditions. Detection and localization of intracystic bacteria and examination of the en- and excystment dynamics is a major challeng...

journal_title：BioTechniques

authors： Lambrecht E,Baré J,Claeys M,Chavatte N,Bert W,Sabbe K,Houf K

更新日期：2015-04-01 00:00:00

abstract：:Naturstoff reagent A (diphenylboric acid 2-aminoethyl ester [DPBA]) has been used historically in plant science to observe polyphenolic pigments, such as flavonoids, whose fluorescence requires enhancement to be visible by microscopy. Flavonoids are common dietary constituents and are the focus of considerable attenti...

journal_title：BioTechniques

authors： Ferrara BT,Thompson EP

更新日期：2019-02-01 00:00:00

abstract：:Much effort has been directed toward the isolation and characterization of homeobox cDNAs from numerous cell types because they encode transcription factors important to many cellular processes, including pattern formation in the embryo, cell growth and cell differentiation. Many novel homeobox cDNAs have been isolate...

journal_title：BioTechniques

authors： Gorski DH,LePage DF,Walsh K

更新日期：1994-05-01 00:00:00