abstract：:Procedures utilizing Chelex 100 chelating resin have been developed for extracting DNA from forensic-type samples for use with the PCR. The procedures are simple, rapid, involve no organic solvents and do not require multiple tube transfers for most types of samples. The extraction of DNA from semen and very small blo...

journal_title：BioTechniques

authors： Walsh PS,Metzger DA,Higuchi R

更新日期：1991-04-01 00:00:00

abstract：:Using a simple electronic circuit, a thermocouple can be connected to a chart recorder to measure the actual temperature inside a PCR tube. This allows accurate inspection of the thermocycle program and comparison between thermoprofiles from different thermocyclers. We found that the recording of temperature cycling e...

journal_title：BioTechniques

authors： Stamm S,Gillo B,Brosius J

更新日期：1991-04-01 00:00:00

abstract：:In this communication we describe the sequential use of standard and low-melting agarose in a single gel slab for the electrophoresis of DNA. This method has the advantages of high resolution and reproducibility characteristic of standard agarose and the ease of manipulation of DNA for direct cloning, sequential diges...

journal_title：BioTechniques

authors： Fotadar U,Shapiro LE,Surks MI

更新日期：1991-02-01 00:00:00

abstract：:A method for the determination of prostaglandin G/H synthase and lipoxygenase activities in tissues was developed and employed with rat gastric mucosa samples. Tissues and microsomes were incubated in a buffer containing nonionic detergent and 1.32 mM arachidonic acid for 10 min. Following extraction with ethyl acetat...

journal_title：BioTechniques

authors： Stein TA,Bailey B,Auguste LJ,Wise L

更新日期：1991-02-01 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：:We report a modification of a liquid hybridization method that serves as a rapid, sensitive alternative to Southern hybridization for the analysis of polymerase chain reaction (PCR) products. An aliquot of the completed PCR is mixed with an internally nested, end-labeled oligonucleotide probe, and one cycle of PCR is ...

journal_title：BioTechniques

authors： Parker JD,Burmer GC

更新日期：1991-01-01 00:00:00

abstract：:We have demonstrated that efficient polymerase chain reaction amplifications from chromosomal DNA can be carried out using whole bacterial cells as the starting material. Cells from the liquid or solid cultures can be used directly, without any pre-treatment, thus eliminating the need for DNA isolation. ...

journal_title：BioTechniques

authors： Joshi AK,Baichwal V,Ames GF

更新日期：1991-01-01 00:00:00

abstract：:Plasmid pUC18rspL is a 3.788-kilobase pair vector, derived from pUC18, pKK232-8 and pHSG664, which identifies promoter-bearing DNA fragments functional in StrA E. coli by activation of a promoterless streptomycin-sensitive gene cartridge (rspL). Expression of the plasmid-borne rspL gene leads to sensitivity dominance ...

journal_title：BioTechniques

authors： Vockley JG,Pène JJ

更新日期：1990-12-01 00:00:00

abstract：:As more and more researchers are examining proteins that are available only in extremely limited quantities, i.e., cellular extracts or genetic engineering products, it is critical to utilize staining methods that maximize sensitivity. The protocol we describe here--double staining of polyacrylamide electrophoresis ge...

journal_title：BioTechniques

authors： Ross M,Peters L

更新日期：1990-11-01 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：: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：:Few studies have focused on the significance of ras protein levels in human malignancy, in part because of the inherent difficulty in quantitation of the ras gene product. We have developed a method for the enzymatic determination of the ras gene product and have used this method for the quantitation of ras gene produ...

journal_title：BioTechniques

authors： Gutheil JC,Mane SM,Bass KA,Lee EJ,Needleman SW

更新日期：1990-08-01 00:00:00

abstract：:A fluorescent in situ DNA hybridization assay employing a biotinylated DNA probe was used to visualize single copies of human immunodeficiency virus (HIV) proviral DNA in the nuclei and metaphase chromosomes of infected cells. In clonal cell lines that contain either one or two copies of proviral DNA, the efficiency o...

journal_title：BioTechniques

authors： Spadoro JP,Payne H,Lee Y,Rosenstraus MJ

更新日期：1990-08-01 00:00:00

abstract：:We describe a simple and inexpensive method of performing sequencing reactions for 24 single-strand M13 DNA clones in microtiter plates. To simplify elevated temperature incubations during sequencing reactions, two heating blocks were designed to accommodate microtiter plates and fit within common laboratory heating m...

journal_title：BioTechniques

authors： Koop BF,Wilson RK,Chen C,Halloran N,Sciammis R,Hood L,Lindelien JW

更新日期：1990-07-01 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：:WE present a rapid procedure based on the polymerase chain reaction for generation of double-stranded DNA templates suitable for in vitro transcription by T3 or T7 RNA polymerase. DNA fragments cloned into a phage promoter vector are amplified together with a flanking promoter to provide functional templates. Extensio...

journal_title：BioTechniques

authors： Weier HU,Rosette C

更新日期：1990-03-01 00:00:00

abstract：:We describe a new real-time fluorescence video microscope design for capturing intensified images of cells containing dual wavelength "ratio" dyes or multiple dyes. The microscope will perform real-time capture of two separate fluorescence emission images simultaneously, improving the time resolution of spatial distri...

journal_title：BioTechniques

authors： Morris SJ

更新日期：1990-03-01 00:00:00

abstract：:A method for the isolation of total cytoplasmic RNA and high molecular weight DNA from the same cells is described. Cells are gently lysed with NP40 in the presence of vanadyl ribonucleoside complex and the nuclei pelleted by centrifugation. RNA is purified by phenol/CHCl3 extraction of the lysate supernatant followed...

journal_title：BioTechniques

authors： Nicolaides NC,Stoeckert CJ Jr

更新日期：1990-02-01 00:00:00

abstract：:Site-specific mutagenesis and directional subcloning were accomplished by using the polymerase chain reaction to generate products that can recombine to form circular DNA. This DNA was transfected into E. coli without phosphorylation of primers, restriction enzyme digestion or ligation. Specifically, the polymerase ch...

journal_title：BioTechniques

authors： Jones DH,Howard BH

更新日期：1990-02-01 00:00:00

abstract：:A rapid and highly sensitive technique (MAPPing: message amplification phenotyping) has been developed to simultaneously analyze the array of messenger RNAs made by small numbers of cells. The technique incorporates a micro-procedure for isolating RNA, reverse transcription of total cellular RNA to produce cDNA, and e...

journal_title：BioTechniques

authors： Brenner CA,Tam AW,Nelson PA,Engleman EG,Suzuki N,Fry KE,Larrick JW

更新日期：1989-11-01 00:00:00

abstract：:The analysis of DNA restriction fragment length polymorphisms by Southern blot hybridization requires that sufficient quantities of high molecular weight genomic DNA be extracted from biological specimens. Prior to analysis, it is necessary to determine the quantity and quality of the extracted DNA. For many applicati...

journal_title：BioTechniques

authors： Waye JS,Presley LA,Budowle B,Shutler GG,Fourney RM

更新日期：1989-09-01 00:00:00

abstract：:We report a straightforward methodology for purification of recombinant proteins by incorporating a short hydrophilic peptide marker segment at their N-termini. A calcium-dependent antibody that reacts primarily with the first three amino acids of this peptide segment was used to affinity purify the fusion proteins in...

journal_title：BioTechniques

authors： Prickett KS,Amberg DC,Hopp TP

更新日期：1989-06-01 00:00:00

abstract：:This report describes a common method of obtaining template DNA from phagemids, phages and plasmids. The strategy is based on the use of the cationic detergent cetyl-trimethylammonium bromide (CTAB) for DNA precipitation. By avoiding phase separation, many manipulation steps are reduced. A time-saving modification to ...

journal_title：BioTechniques

authors： Del Sal G,Manfioletti G,Schneider C

更新日期：1989-05-01 00:00:00

abstract：:The microcomputer-based image analysis system IB-1000 (developed by Indiana Biotech, Highland, IN) for two-dimensional electrophoresis gels has been described previously (9). It allows the user to compare protein spots between two profiles and identify those spots that are commonly shared in both profiles. This report...

journal_title：BioTechniques

authors： Lee C,Sherwood ER,Sensibar JA,Berg LA,Chen YC,Tseng CC

更新日期：1989-04-01 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 head-to-tail trimer of the SV40 Bcl I-Bam H1 DNA fragment, specifying polyadenylation of RNA transcripts, was cloned as a cassette flanked by multiple restriction sites. Insertion of the trimer into several expression vectors efficiently prevented spurious expression of reporter genes resulting from transcriptional ...

journal_title：BioTechniques

authors： Maxwell IH,Harrison GS,Wood WM,Maxwell F

更新日期：1989-03-01 00:00:00

abstract：:Several techniques were evaluated for the quantitation of the total protein content of an IgG2a monoclonal antibody, KS1/4, and its deacetylvinblastine (DAVLB) conjugate. The UV assay is rapid, but it requires an extensive calibration of the response factor, and impurities may cause a high bias. Amino acid analysis (A...

journal_title：BioTechniques

authors： Rickard EC,Sittampalam GS,Clodfelter DK

更新日期：1988-11-01 00:00:00

abstract：:To establish a nonradioactive method for demonstrating HPV DNA in routinely treated smears of the uterine cervix (alcohol fixation, staining according to Papanicolaou, preservation), in situ hybridizations were carried out in HeLa and SiHa cells grown on slides. After detailed investigations, the sensitivity and speci...

journal_title：BioTechniques

authors： Heiles HB,Genersch E,Kessler C,Neumann R,Eggers HJ

更新日期：1988-11-01 00:00:00

abstract：:We report here a system for automation of the dideoxynucleotide DNA sequencing method. The system consists of a Beckman Biomek 1000 robotic workstation which has been modified by the addition of a thin heater/cooler block directly on the instrument table. The heater/cooler block, which is regulated by a user-specified...

journal_title：BioTechniques

authors： Wilson RK,Yuen AS,Clark SM,Spence C,Arakelian P,Hood LE

更新日期：1988-09-01 00:00:00

abstract：:Transfection experiments involving mixed cell cultures pose special problems because of differential uptake and expression of DNA by different cell types. In order to evaluate the expression of transfected DNA in primary cultures of embryonic myocardial cells, we have used immunofluorescence microscopy to analyze expr...

journal_title：BioTechniques

authors： Antin PB,Mar JH,Ordahl CP

更新日期：1988-07-01 00:00:00

abstract：:Patents play an increasingly important role in the dissemination of information in many fast moving fields such as biotechnology and semiconductors. Quite a few new developments are introduced as patents, and only later, if at all, do they find their way into the scientific literature. In spite of this, patents lack w...

journal_title：BioTechniques

authors： Koniarek JP,Coleman KD

更新日期：1988-02-01 00:00:00