Optimized multiplex IgH/ras PCR: a tool for quantitative monitoring of B-lymphoproliferative disorders.

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) as a control gene. Serial dilutions of stored diagnostic DNAs are examined together in the same PCR at a sub-plateau phase and, after analysis by densitometry, the amount of CDR3 product is related to the ras product. An increase of this ratio at comparable amounts of DNA is viewed as an increase of malignant cells. This endpoint PCR quantifying approach appears to be applicable in monitoring B-lymphoproliferative disorders as was shown to be true in B-cell non-Hodgkin's lymphoma and may provide information on disease activity and treatment outcome.

journal_name

Biotechniques

journal_title

BioTechniques

authors

Slavícková A,Ullmannová V,Klener P

doi

10.2144/00284st08

subject

Has Abstract

pub_date

2000-04-01 00:00:00

pages

716-8, 720-1

issue

4

eissn

0736-6205

issn

1940-9818

journal_volume

28

pub_type

杂志文章
  • Pilot-scale production of murine monoclonal antibodies in agitated, ceramic-matrix or hollow-fiber cell culture systems.

    abstract::The purpose of this research was to compare three bioreactor systems for the pilot-scale production of monoclonal antibodies (MAbs). We needed to produce gram quantities of murine MAbs against human prostatic acid phosphatase for use in fragmentation, radiolabeling and in vivo radio-imaging studies. The stable hybrido...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:

    authors: Kurkela R,Fraune E,Vihko P

    更新日期:1993-10-01 00:00:00

  • Large-scale supercoiled plasmid preparation by acidic phenol extraction.

    abstract::A novel method for large-scale plasmid preparation is described. Crude extracts are subjected to acidic phenol extraction to remove any contaminants present in the aqueous phase. The supercoiled plasmid DNA, which preferentially remains in the organic phase and inter-phase, is extracted back into the aqueous phase wit...

    journal_title:BioTechniques

    pub_type:

    doi:

    authors: Wang Z,Rossman TG

    更新日期:1994-03-01 00:00:00

  • Use of an ALFexpress DNA sequencer to analyze protein-nucleic acid interactions by band shift assay.

    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

    pub_type: 杂志文章

    doi:10.2144/01305rr03

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

    更新日期:2001-05-01 00:00:00

  • Analysis of mixed human/microbial DNA samples: a validation study of two PCR AMP-FLP typing methods.

    abstract::The reliability of the PCR technique used to type two human variable number tandem repeats, that is, 3' to apolipoprotein B gene and locus D17S30, was examined using DNA samples of mixed human and microbial origin. Mixtures of human and microbial DNA were amplified, choosing microbes found commonly in the vagina. Tota...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:

    authors: Lienert K,Fowler JC

    更新日期:1992-08-01 00:00:00

  • Low-cost image processing system for analyzing molecular RNA fingerprints.

    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

    pub_type: 杂志文章

    doi:

    authors: Mitchell BW,Palmieri S

    更新日期:1990-10-01 00:00:00

  • Quantitative RT-PCR: pitfalls and potential.

    abstract::Reverse transcription PCR (RT-PCR) represents a sensitive and powerful tool for analyzing RNA. While it has tremendous potential for quantitative applications, a comprehensive knowledge of its technical aspects is required. Successful quantitative RT-PCR involves correction for experimental variations in individual RT...

    journal_title:BioTechniques

    pub_type: 杂志文章,评审

    doi:10.2144/99261rv01

    authors: Freeman WM,Walker SJ,Vrana KE

    更新日期:1999-01-01 00:00:00

  • SNPCEQer: detecting SNPs in sequences generated by the Beckman CEQ2000 DNA Analysis System.

    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

    pub_type:

    doi:10.2144/02334st05

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

    更新日期:2002-10-01 00:00:00

  • Short technical reports. Modification of the TRI reagent procedure for isolation of RNA from polysaccharide- and proteoglycan-rich sources.

    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

    pub_type: 杂志文章

    doi:

    authors: Chomczynski P,Mackey K

    更新日期:1995-12-01 00:00:00

  • Message amplification phenotyping (MAPPing): a technique to simultaneously measure multiple mRNAs from small numbers of cells.

    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

    pub_type: 杂志文章

    doi:

    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

  • Formalin removal from archival tissue by critical point drying.

    abstract::The extraction of high-quality nucleic acid may be problematic in formalin-fixed tissues because of cross-linking between proteins and DNA. Old fixed tissue specimens do produce fragmented DNA (<1.2 kb), which is only used for PCR amplification. Here we show that high molecular weight DNA (>194 kb) can be successfully...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/02333rr03

    authors: Fang SG,Wan QH,Fujihara N

    更新日期:2002-09-01 00:00:00

  • Rapid identification of transformed wheat using a half-seed PCR assay.

    abstract::A simple, nondestructive PCR-based screening method has been developed for identifying putative transgenic soft white winter wheat (Triticum aestivum L.) carrying the coat protein gene of wheat streak mosaic virus. Removal of the endosperm end of individual seed provided sufficient material for DNA extraction and PCR....

    journal_title:BioTechniques

    pub_type:

    doi:10.2144/02323st09

    authors: McCarthy PL,Hansen JL,Zemetra RS,Berger PH

    更新日期:2002-03-01 00:00:00

  • Creating random mutagenesis libraries using megaprimer PCR of whole plasmid.

    abstract::The conventional method for cloning a DNA fragment is to insert it into a vector and ligate it. Although this method is commonly used, it is labor intensive because the ratio and concentrations of the DNA insert and the vector need optimizing. Even then, the resultant library is often plagued with unwanted plasmids th...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/02335st03

    authors: Miyazaki K,Takenouchi M

    更新日期:2002-11-01 00:00:00

  • An improved Huffman coding method for archiving text, images, and music characters in DNA.

    abstract::An improved Huffman coding method for information storage in DNA is described. The method entails the utilization of modified unambiguous base assignment that enables efficient coding of characters. A plasmid-based library with efficient and reliable information retrieval and assembly with uniquely designed primers is...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/000113218

    authors: Ailenberg M,Rotstein O

    更新日期:2009-09-01 00:00:00

  • Adenovirus-mediated transfection of cultured cells.

    abstract::We describe here a simple and efficient transfection method for transient expression of cloned genes in cell lines and primary cultured cells. The method involves the use of DEAE-dextran to target DNA to the cellular endocytotic pathway and the use of a human adenovirus to ensure efficient lysis of endosomal vesicles....

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:

    authors: Forsayeth JR,Garcia PD

    更新日期:1994-08-01 00:00:00

  • Mammalian two-hybrid system: a complementary approach to the yeast two-hybrid system.

    abstract::Here we demonstrate the use of a mammalian two-hybrid system to study protein-protein interactions. Like the yeast two-hybrid system, this is a genetic, in vivo assay based on the reconstitution of the function of a transcriptional activator. In this system, one protein of interest is expressed as a fusion to the Gal4...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/97222pf02

    authors: Luo Y,Batalao A,Zhou H,Zhu L

    更新日期:1997-02-01 00:00:00

  • Single copies of HIV proviral DNA detected by fluorescent in situ hybridization.

    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

    pub_type: 杂志文章

    doi:

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

    更新日期:1990-08-01 00:00:00

  • DIRECTING DRUG EVOLUTION.

    abstract::Directed evolution is poised to change small molecule discovery and provide greater access to "drug space." Sarah Webb looks into the evolving drug discovery landscape. ...

    journal_title:BioTechniques

    pub_type: 新闻

    doi:10.2144/000114616

    authors: Webb S Ph D

    更新日期:2017-12-01 00:00:00

  • Absolute quantitation of cancer-related proteins using an MS-based peptide chip.

    abstract::New technologies are needed that can diagnose cancer more rapidly and accurately. These technologies must also have the ability to identify the particular cellular abnormalities contributing to the malignancy, thus directing the appropriate treatments. Such technologies should permit absolute quantitation of specific ...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/05386su01

    authors: Warren EN,Jiang J,Parker CE,Borchers CH

    更新日期:2005-06-01 00:00:00

  • Evaluation of DNA degradation and establishment of a degradation analysis model for Lepidoptera specimens.

    abstract::Millions of museum specimens are integral to biodiversity studies; however, DNA degradation may limit the ability to obtain DNA sequences. In this study, a degradation analysis model for Lepidoptera specimens was established. Based on this model, we revealed the characteristics of DNA fragment distribution caused by e...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/btn-2019-0166

    authors: Xu Y,Ren X,Wang H,Wang M,Li G

    更新日期:2020-03-01 00:00:00

  • Surrogate marker for predicting the virus binding of urogenital cancer cells during adenovirus-based gene therapy.

    abstract::Virus uptake is the first rate-limiting step for the successful gene delivery of any virus-based gene therapy. For adenovirus-based gene therapy, the expression levels of the adenovirus receptor--coxsackievirus and adenovirus receptor (CAR)--play an important role in dictating gene delivery. We have observed a wide sp...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/03351dd02

    authors: Lai YJ,Pong RC,McConnell JD,Hsieh JT

    更新日期:2003-07-01 00:00:00

  • Mobility-shift analysis with microfluidics chips.

    abstract::Electrophoretic mobility shift analysis (EMSA) is a well-characterized and widely used technique for the analysis of proten-DNA interaction and the analysis of transcription factor combinatorics. Currently implemented EMSA generally involves the time-consuming use of radiolabeled DNA and polyacrylamide gel electrophor...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/03353rr01

    authors: Clark J,Shevchuk T,Swiderski PM,Dabur R,Crocitto LE,Buryanov YI,Smith SS

    更新日期:2003-09-01 00:00:00

  • A device for stereotaxic viral delivery into the brains of neonatal mice.

    abstract::The increasing interest in manipulating neural circuits in developing brains has created a demand for reliable and accurate methods for delivering viruses to newborn mice. Here we describe a novel 3D-printed mouse neonatal stereotaxic adaptor for intracerebral viral injection that provides enhanced precision and relia...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/btn-2020-0050

    authors: Olivetti PR,Lacefield CO,Kellendonk C

    更新日期:2020-10-01 00:00:00

  • Graphene veils: A versatile surface chemistry for sensors.

    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

    pub_type: 杂志文章

    doi:10.2144/000114188

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

    更新日期:2014-07-01 00:00:00

  • pGATA: a positive selection vector based on the toxicity of the transcription factor GATA-1 to bacteria.

    abstract::The transcription factor GATA-1 is a zinc finger DNA-binding protein essential for the development of red blood cells. When we expressed different regions of the zinc finger domain in bacteria using an isopropyl-beta-D-thiogalactoside (IPTG) inducible system, growth of bacteria harboring the active DNA-binding domain ...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/19962004684

    authors: Trudel P,Provost S,Massie B,Chartrand P,Wall L

    更新日期:1996-04-01 00:00:00

  • A TB40/E-derived human cytomegalovirus genome with an intact US-gene region and a self-excisable BAC cassette for immunological research.

    abstract::For immunological research on the human cytomegalovirus (HCMV), a virus that combines the broad cell tropism of clinical isolates, efficient replication in cell culture, the complete set of MHC-I modulator genes, and suitability for genetic engineering is desired. Here, we aimed to generate a genetically complete deri...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/000114606

    authors: Sampaio KL,Weyell A,Subramanian N,Wu Z,Sinzger C

    更新日期:2017-11-01 00:00:00

  • Use of signal-distinguishable probes in differential or sequential autoradiography in hybridization analysis.

    abstract::To avoid the time-consuming reprobing process in hybridization analysis, signal-distinguishable probes (32P, 35S or antigenic hapten-labeled DNA) can be added to the same hybridization mixture. After hybridization, an unambiguous result can be obtained by differential or sequential autoradiography. ...

    journal_title:BioTechniques

    pub_type:

    doi:

    authors: Au LC,Chang KJ,Shih CM,Teh GW

    更新日期:1994-04-01 00:00:00

  • Rapid high resolution western blotting: from gel to image in a single day.

    abstract::A streamlined protocol is described that allows high sensitivity antigen detection by Western blotting in a single day. The choice of membrane blotting matrix, as well as blocking reagents, has been optimized in order to allow rapid development of the blot with chemiluminescent reagents. The entire process, from gel t...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:

    authors: Klapper A,MacKay B,Resh MD

    更新日期:1992-05-01 00:00:00

  • Accurate and efficient data processing for quantitative real-time PCR using a tripartite plant virus as a model.

    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

    pub_type: 杂志文章

    doi:10.2144/000112750

    authors: Feng J,Zeng R,Chen J

    更新日期:2008-06-01 00:00:00

  • A microbiological assay for the quantitative determination of glutathione.

    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

    pub_type: 杂志文章

    doi:10.2144/96215st07

    authors: Schmidt M,Grey M,Brendel M

    更新日期:1996-11-01 00:00:00

  • Method for isolation of PCR-ready genomic DNA from zebrafish tissues.

    abstract::Here we describe a method for the isolation of PCR-ready genomic DNA from various zebrafish tissues that is based on a previously published murine protocol. The DNA solutions are of sufficient quality to allow PCR detection of transgenes from all commonly used zebrafish tissues. In sperm, transgene amplification was s...

    journal_title:BioTechniques

    pub_type: 杂志文章

    doi:10.2144/000112619

    authors: Meeker ND,Hutchinson SA,Ho L,Trede NS

    更新日期:2007-11-01 00:00:00