A new outlier removal approach for cDNA microarray normalization.

abstract：:Changes in cardiac structure that depart from normal have generally been termed "remodeling". Assessment of ventricular remodeling at the cellular level should include measurement of myocyte dimensions. A well-established and reliable method to assess myocyte remodeling uses isolated cells and the Coulter Counter/Chan...

journal_title：BioTechniques

authors： Said S,Tamura T,Gerdes AM

更新日期：1998-09-01 00:00:00

abstract：:A modification of the TRI Reagent procedure has been elaborated for isolation of RNA from polysaccharide- and proteoglycan-rich material. In the modified procedure, RNA is precipitated from the aqueous phase by the combined action of isopropanol and a high-salt concentration. Under these conditions, RNA is effectively...

journal_title：BioTechniques

authors： Chomczynski P,Mackey K

更新日期：1995-12-01 00:00:00

abstract：:The use of quantitative PCR is recommended to monitor the level of residual hematological malignancies. The proposed multiplex IgH/ras PCR uses a co-amplification of the clonal CDR3 rearrangement of the immunoglobulin heavy chain gene (IgH) as a disease marker and a segment of the Hras 1 gene containing codon 61 (ras)...

journal_title：BioTechniques

authors： Slavícková A,Ullmannová V,Klener P

更新日期：2000-04-01 00:00:00

abstract：:New cloning vectors have been developed with features to enhance quick allelic exchange in gram-negative bacteria. The conditionally replicative R6K and transfer origins facilitate conjugation and chromosomal integration into a variety of bacterial species, whereas the sacB gene provides counterselection for allelic e...

journal_title：BioTechniques

authors： Mejia-Santana A,Lloyd CJ,Klose KE

更新日期：2021-01-25 00:00:00

abstract：:The construction and the testing of two lambda phage vectors are described that greatly simplify the tasks of mapping genomic DNA and making replacement-type gene-targeting vectors for mammalian cells from a library of isogenic genomic DNA. The first vector, lambda PS, accommodates up to 20 kb and allows inserts to be...

journal_title：BioTechniques

authors： Nehls M,Messerle M,Sirulnik A,Smith AJ,Boehm T

更新日期：1994-10-01 00:00:00

abstract：:Based on the requirement of a glutathione-deficient mutant strain of Saccharomyces cerevisiae to take up external glutathione for growth on synthetic media, a simple agar diffusion test for quantitative detection of total glutathione from various sources was established. Glutathione concentrations can be reliably dete...

journal_title：BioTechniques

authors： Schmidt M,Grey M,Brendel M

更新日期：1996-11-01 00:00:00

abstract：:We describe a simple and efficient technique that facilitates the amplification of specific mRNA for cloning and sequencing purposes. An mRNA bound to a small piece of membrane filter is used as a template to synthesize complementary DNA. The product of this reaction is then transferred to a new tube and amplified usi...

journal_title：BioTechniques

authors： Ruiz A,Bok D

更新日期：1993-11-01 00:00:00

abstract：:RNA interference (RNAi) is a recent advance that provides the possibility to reduce the expression of specific target genes in cultured mammalian cells with potential applications on a genome-wide scale. However, to achieve this, robust methodologies that allow automated and efficient delivery of small interfering RNA...

journal_title：BioTechniques

authors： Erfle H,Simpson JC,Bastiaens PI,Pepperkok R

更新日期：2004-09-01 00:00:00

abstract：:As a strategy to purify recombinant murine Interleukin (IL)-18, we cloned the mature coding region of this protein into the pFLAG-1 expression system. The intent was to use the FLAG peptide "tag" as an amino terminal addition to IL-18 so that purification of this fusion protein (FLAG-IL-18) on anti-FLAG antibody affin...

journal_title：BioTechniques

authors： Elhofy A,Bost KL

更新日期：1998-09-01 00:00:00

abstract：:To measure mutation load or to detect minimal residual disease, a robust method for identifying one mutant allele in the range of 10(6)-10(9) wild-type alleles would be advantageous. Herein, we present evidence that pyrophosphorolysis-activated polymerization (PAP) has the potential to provide a highly specific and ro...

journal_title：BioTechniques

authors： Liu Q,Sommer SS

更新日期：2000-11-01 00:00:00

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：:Neuronal death can be induced by DNA-damaging agents and occurs by apoptosis involving a specific signal-transduction pathway. However, to our knowledge, methods for the quantitative determination of DNA damage in individual neurons have not yet been described. Here we optimize the single-cell gel electrophoresis (SCG...

journal_title：BioTechniques

authors： Morris EJ,Dreixler JC,Cheng KY,Wilson PM,Gin RM,Geller HM

更新日期：1999-02-01 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：:Identification of nucleotides used for RNA chain initiation or for contacting DNA binding proteins is basic to our understanding of gene regulation. Normally, a radioactive primer is used to copy RNA or DNA. The polymerase extension stops at free ends of mRNA (as in promoter mapping) or at the position of template cle...

journal_title：BioTechniques

authors： Fekete RA,Miller MJ,Chattoraj DK

更新日期：2003-07-01 00:00:00

abstract：:Plasmid DNA can be purified in 20 min using a three-step CsCl gradient in a vertical tube rotor. Ultracentrifugation with discontinuous gradients yields DNA with purity comparable to that obtained with longer isopycnic separations and with columns. ...

journal_title：BioTechniques

authors： Dorin M,Bornecque CA

更新日期：1995-01-01 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：:Currently, in real-time PCR, one often has to choose between using a sequence-specific probe and a nonspecific double-stranded DNA (dsDNA) binding dye for the detection of amplified DNA products. The sequence-specific probe has the advantage that it only detects the targeted product, while the nonspecific dye has the ...

journal_title：BioTechniques

authors： Lind K,Ståhlberg A,Zoric N,Kubista M

更新日期：2006-03-01 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 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：:Thymidine kinase 1 (TK1) is an enzyme involved in DNA precursor synthesis that has been used as a biomarker for prognosis and monitoring of different malignancies. In this study, we compared two immunoassays for measuring TK1 protein concentrations: the TK 210 ELISA (AroCell AB) and TK1 ELISA from Abcam. Overall, the ...

journal_title：BioTechniques

authors： Kumar JK,Holmgren S,Levedahl KH,Höglund M,Venge P,Eriksson S

更新日期：2020-06-01 00:00:00

abstract：:Substantial technological advances in proteomics and related computational science have been made in the past few years. These advances overcome in part the complexity and heterogeneity of the human proteome, permitting quantitative analysis and identification of protein changes associated with tumor development. Here...

journal_title：BioTechniques

authors： Faca V,Krasnoselsky A,Hanash S

更新日期：2007-09-01 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：:We describe a novel assay format for the Gal4-based yeast two-hybrid-system, in which the readout from three different reporter genes is measured sequentially in a single microplate. Activation of the URA3, MEL1, and lacZ reporters in response to a protein-protein interaction is monitored by measuring sequentially: (i...

journal_title：BioTechniques

authors： Evans DR,Swirsding KA,Taillon BE,Simons JF

更新日期：2004-11-01 00:00:00

abstract：:Real-time PCR is becoming a preferred method for quantification of minute amounts of nucleic acids. To achieve the full potential of this technique, accurate and convenient models for post-PCR data analysis are required. In this study, three different models were chosen to quantify the definitive copy numbers of Cucum...

journal_title：BioTechniques

authors： Feng J,Zeng R,Chen J

更新日期：2008-06-01 00:00:00

abstract：:Conformation-sensitive gel electrophoresis (CSGE) has been introduced as the most reliable method for the screening of large and multi-exon genes because of its simplicity, sensitivity and specificity. Based on heteroduplex formation and with the use of mildly denaturing solvents, it allows detection of single-base mu...

journal_title：BioTechniques

authors： Blesa JR,Hernández-Yago J

更新日期：2000-05-01 00:00:00

abstract：:Phage display holds a key position in the use of combinatorial library approaches for the purpose of protein engineering and discovery. However, modifying the pIII protein of the phage can severely and negatively influence the infectiousness of the phage particle. This concern is particularly relevant when large pIII ...

journal_title：BioTechniques

authors： Løset GÅ,Kristinsson SG,Sandlie I

更新日期：2008-04-01 00:00:00

abstract：:A nascent protein remains in a complex with its ribosome and mRNA if the stop codon is deleted from the mRNA. In the same manner, mRNA forms a stable complex with DNA if the transcription termination is blocked. In principle, if both mRNA translation and DNA transcription termination are prevented, the protein should ...

journal_title：BioTechniques

authors： Liu S,Shivashankar GV,Sano T,Libchaber A

更新日期：2000-10-01 00:00:00

abstract：:Treatment with 2 mM CuSO4 was used to induce a Drosophila melanogaster metallothionein (Mtn) promoter that had been cloned into a recombinant baculovirus. Careful study revealed that the Mtn promoter functioned as an inducible, if somewhat "leaky" promoter within the context of baculovirus-infected cells. In the proce...

journal_title：BioTechniques

authors： Lanier LM,Storm K,Shafaie A,Volkman LE

更新日期：1997-10-01 00:00:00

abstract：:There is an enormous initiative to establish the genetic basis for disorders of brain function. Unfortunately, genetic intervention is not accomplished easily in the nervous system. One strategy is to engineer and deliver to neurons specialized viral vectors that carry a gene (or genes) of interest, thereby exploiting...

journal_title：BioTechniques

authors： Neve RL,Neve KA,Nestler EJ,Carlezon WA Jr

更新日期：2005-09-01 00:00:00

abstract：:Polyethyleneimine (PEI) is a flocculent that is widely used in the downstream purification of monoclonal antibodies. It is an in-process residual that is carried through the drug purification process and strongly inhibits residual DNA quantitation by real-time quantitative PCR assay. Very high sample dilutions (e.g., ...

journal_title：BioTechniques

authors： Zhang SM,Roberts M,Jones M,Zeitler J,Kilby G,Liu A,White JR

更新日期：2020-06-01 00:00:00