Colorimetric detection of loop-mediated isothermal amplification reaction by using hydroxy naphthol blue.

abstract：:Gel retardation analysis, or band shift assay, is technically the simplest method to investigate protein-nucleic acid interactions. In this report, we describe a nonradioactive band shift assay using a fluorescent DNA target and an ALFexpress automatic DNA sequencer in place of the current method that utilizes radioac...

journal_title：BioTechniques

authors： Filée P,Delmarcelle M,Thamm I,Joris B

更新日期：2001-05-01 00:00:00

abstract：:A rapid and inexpensive method for isolating bacterial DNA for use in PCR is described. The method is based on the guanidinium thiocyanate (GuSCN)-lysis method of Boom et al. (J. Clin. Microbiol. 28:495-503, 1990) and enables a multiple of 96 samples to be prepared in only one hour. We use Multiscreen plates and a vac...

journal_title：BioTechniques

authors： Reek FH,Smits MA,Kamp EM,Smith HE

更新日期：1995-08-01 00:00:00

abstract：:Single-stranded long oligonucleotide-based (50- to 70-mer) microarrays offer several advantages over conventional cDNA microarrays. These include the easy preparation of the probes, low cost of array production, and low cross-contamination during probe handling. However, the application of oligonucleotide microarrays ...

journal_title：BioTechniques

authors： Kaposi-Novak P,Lee JS,Mikaelyan A,Patel V,Thorgeirsson SS

更新日期：2004-10-01 00:00:00

abstract：:We observed the presence of contaminating NADH oxidation activity in maltose binding protein (MBP) fusion proteins expressed in Escherichia coli and purified using conventional amylose resin-based affinity chromatography. This contaminating NADH oxidation activity was detectable with at least four different enzymes fr...

journal_title：BioTechniques

authors： Guo F,Zhu G

更新日期：2012-04-01 00:00:00

abstract：:Genome sequencing projects are either based on whole genome shotgun (WGS) or on a BAC-by-BAC strategy. Although WGS is in most cases the preferred choice, sometimes the BAC-by-BAC approach may be better because it requires a much simpler assembly process. Furthermore, when the study is limited to specific regions of t...

journal_title：BioTechniques

authors： Todesco S,Campagna D,Levorin F,D'Angelo M,Schiavon R,Valle G,Vezzi A

更新日期：2008-01-01 00:00:00

abstract：:A dot immunoblotting technique has been developed to estimate the relative expression levels of tagged recombinant human proteins in mammalian cell culture media. Variations in sample denaturation, blocking agents and membrane composition and treatment were used to optimize the signal-to-noise ratio of the defined pro...

journal_title：BioTechniques

authors： Ellsworth JL,Hamacher N,Bagwell N,West JW

更新日期：2000-05-01 00:00:00

abstract：:Xer-cise is a technique using antibiotic resistance cassettes flanked by dif sites allowing spontaneous and accurate excision from bacterial chromosomes with a high frequency through the action of the cellular recombinase XerCD. Here, we report a significant improvement of Xer-cise in Mycobacteria. Zeocin resistance c...

journal_title：BioTechniques

authors： Boudehen YM,Wallat M,Rousseau P,Neyrolles O,Gutierrez C

更新日期：2020-02-01 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：:Chitin from crustacean shells has often been used to isolate and purify plant lectins that have an affinity for poly-N-acetylglucosamine (poly-GlcNAc). When we used washed chitin from crab shells as an affinity medium to isolate a lectin from Pinus strobus L. (eastern white pine) ovules, we found that a substance havi...

journal_title：BioTechniques

authors： Whitmore FA

更新日期：1992-02-01 00:00:00

abstract：:Caspase-3 is a key effector caspase that is activated in both extrinsic and intrinsic pathways of apoptosis. Available fluorescent sensors for caspase-3 activity operate in relatively short wavelength regions and are nonoptimal for multiparameter microscopy and whole-body imaging. In the present work, we developed new...

journal_title：BioTechniques

authors： Zlobovskaya OA,Sergeeva TF,Shirmanova MV,Dudenkova VV,Sharonov GV,Zagaynova EV,Lukyanov KA

更新日期：2016-02-01 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：:Real-time or quantitative loop-mediated isothermal amplification (qLAMP) is a promising technique for the accurate detection of pathogens in organisms and the environment. Here we present a comparative study of the performance of six fluorescent intercalating dyes-SYTO-9, SYTO-13, SYTO-82, SYBR Green I, SYBR Gold, Eva...

journal_title：BioTechniques

authors： Oscorbin IP,Belousova EA,Zakabunin AI,Boyarskikh UA,Filipenko ML

更新日期：2016-07-01 00:00:00

abstract：:High affinity aptamer-based biomarker discovery has the advantage of simultaneously discovering an aptamer affinity reagent and its target biomarker protein. Here, we demonstrate a morphology-based tissue aptamer selection method that enables us to use tissue sections from individual patients and identify high-affinit...

journal_title：BioTechniques

authors： Wang H,Li X,Volk DE,Lokesh GL,Elizondo-Riojas MA,Li L,Nick AM,Sood AK,Rosenblatt KP,Gorenstein DG

更新日期：2016-11-01 00:00:00

abstract：:We present a simple method for sequential chemiluminescent detections of two different DNA loci on a single Southern blot. First, an enzyme-linked DNA probe for a unique sequence is detected with a horse-radish peroxidase (HRP) substrate followed by the detection of another enzyme-linked DNA probe for a different uniq...

journal_title：BioTechniques

authors： Reddy LV,DeSilva R,Handley RS,Schaap AP,Akhavan-Tafti H

更新日期：1999-04-01 00:00:00

abstract：:The technique described here is a fast and simple method of extracting chloroplast DNA (cpDNA). It overcomes the need for differential centrifugation using density gradients. The leaves do not have to be kept in the dark and lyophilized before extraction, but lyophilization is still possible. The chloroplasts are spec...

journal_title：BioTechniques

authors： Mariac C,Trouslot P,Poteaux C,Bezançon G,Renno JF

更新日期：2000-01-01 00:00:00

abstract：:Thin spun-coat films (~4 nm thick) of graphene oxide (GO) constitute a versatile surface chemistry compatible with a broad range of technologically important sensor materials. Countless publications are dedicated to the nuances of surface chemistries that have been developed for sensors, with almost every material hav...

journal_title：BioTechniques

authors： Mulvaney SP,Stine R,Long NC,Tamanaha CR,Sheehan PE

更新日期：2014-07-01 00:00:00

abstract：:At many research institutions, lab space is more valuable than gold. Developers are taking note by designing smaller instruments with enhanced capabilities. Nathan Blow looks inside today's tiny lab. ...

journal_title：BioTechniques

authors： Blow NS Ph D

更新日期：2017-01-01 00:00:00

abstract：:Multiplex Manager 1.0 is a user-friendly cross-platform program that designs efficient combinations of existing genetic marker loci into multiplex polymerase chain reactions and optimizes using prior marker information. The program has the flexibility to solve two design problems: combining all markers into the smalle...

journal_title：BioTechniques

authors： Holleley CE,Geerts PG

更新日期：2009-06-01 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：:MicroRNAs (miRNAs) are ~22-nucleotide-long small non-coding RNAs that regulate the expression of protein-coding genes by base pairing to partially complementary target sites, preferentially located in the 3´ untranslated region (UTR) of target mRNAs. The expression and function of miRNAs have been extensively studied ...

journal_title：BioTechniques

authors： Persson H,Søkilde R,Pirona AC,Rovira C

更新日期：2017-08-01 00:00:00

abstract：:DNA-binding proteins can be purified by their affinity for probes with specific sequences that are tethered to magnetic beads. A restriction site at the end of the tether allows release of probe with factor still on it. This probe with bound factor can be bandshifted directly to compare to bandshifts in whole extracts...

journal_title：BioTechniques

authors： Ren L,Chen H,Sternberg EA

更新日期：1994-05-01 00:00:00

abstract：:Current gene synthesis methods often incorporate a PCR amplification step in order to yield final material sufficient for resolution from multiple off-products. These amplification steps can cause stochastic sampling effects that propagate errors in gene synthesis or decrease variability when applied to the constructi...

journal_title：BioTechniques

authors： Lee ZB,Firnhaber C,Clarke J,DeDecker BS

更新日期：2015-09-01 00:00:00

abstract：:Replacement of colorimetric substrates in Western blots by chemiluminescent reagents enhances the sensitivity of detection by more than an order of magnitude. Protein levels beyond the detection limit of colorimetric substrates can be consistently detected without modifying pre-existing laboratory protocols. ...

journal_title：BioTechniques

authors： Sandhu GS,Eckloff BW,Kline BC

更新日期：1991-07-01 00:00:00

abstract：:A method for cloning single-stranded oligonucleotides in a plasmid vector has been developed. The method relies on ligation of the oligonucleotide into suitable restriction enzyme sites of the cloning vector such that the site at the 5' end has a 5' overhang [for example, a Bgl II site (A decreases GATCT)], and the si...

journal_title：BioTechniques

authors： Mounts P,Wu TC,Peden K

更新日期：1989-04-01 00:00:00

abstract：:A complete image digitizing and processing system is described for capturing, enhancing and analyzing molecular fingerprints. The low-cost, high-resolution system features a Motorola 68000 processor, multi-tasking, a separate video coprocessor, and color or gray scale processing. Thousands of manipulations are possibl...

journal_title：BioTechniques

authors： Mitchell BW,Palmieri S

更新日期：1990-10-01 00:00:00

abstract：:Here we describe our progress in the development of the technology of DNA sequencing by primer walking based on the modular primer approach, which eliminates the primer synthesis bottleneck. Modular primers are assembled from 5-mers, 6-mers, or 7-mers selected from a presynthesized library of as few as 1000 oligonucle...

journal_title：BioTechniques

authors： Kotler L,Sobolev I,Ulanovsky L

更新日期：1994-09-01 00:00:00

abstract：:Next-generation sequencing (NGS) of multigene panels performed for genetic clinical diagnostics requires 100% coverage of all targeted genes. In the genetic diagnostics laboratory, coverage gaps are typically filled with Sanger sequencing after NGS data are collected and analyzed. Libraries prepared using the hybridiz...

journal_title：BioTechniques

authors： Coonrod EM,Durtschi JD,VanSant Webb C,Voelkerding KV,Kumánovics A

更新日期：2014-10-01 00:00:00

abstract：:Manipulating gene expression in primary neurons has been a goal for many scientists for over 20 years. Vertebrate central nervous system neurons are classically difficult to transfect. Most lipid reagents are inefficient and toxic to the cells, and time-consuming methods such as viral infections are often required to ...

journal_title：BioTechniques

authors： Buchser WJ,Pardinas JR,Shi Y,Bixby JL,Lemmon VP

更新日期：2006-11-01 00:00:00

abstract：:We have developed a rapid [3H]colchicine competition-binding scintillation proximity assay (SPA) to evaluate antimitotic compounds that bind to the colchicine-binding site on tubulin. The premise of our assay is that compounds will compete with radiolabeled colchicine for the tubulin-binding domain. Biotin-labeled tub...

journal_title：BioTechniques

authors： Tahir SK,Kovar P,Rosenberg SH,Ng SC

更新日期：2000-07-01 00:00:00

abstract：:The colony formation assay (CFA) is the gold standard for measuring the effects of cytotoxic agents on cancer cells in vitro; however, in its traditional 6-well format, it is a time-consuming assay, particularly when evaluating combination therapies. In the interest of increased efficiency, the 6-well CFA was converte...

journal_title：BioTechniques

authors： Katz D,Ito E,Lau KS,Mocanu JD,Bastianutto C,Schimmer AD,Liu FF

更新日期：2008-02-01 00:00:00