Abstract：
:Few studies have compared the quantification of mRNA by DNA microarray to the results obtained by reverse transcription PCR (RT-PCR). In this study, mRNA was collected from the healing femoral fracture callus of adult and juvenile rats at various times after fracture. Ten samples were measured by both methods for 26 genes. For RT-PCR, mRNA was reverse transcribed, amplified, electrophoresed, blotted, and probed with 32P-labeled internal oligonucleotides, which were quantified. For DNA microarray, the mRNA was processed to biotin-labeled cRNA, hybridized to 10 Affymetrix Rat U34A microarrays, and quantified. Correlation coefficients (r) for each gene for the agreement between RT-PCR and microarray ranged from -0.48 to +0.93. This variation made the interpretation gene-specific. Genes with moderate expression levels gave the highest r values. Increased numbers of absent calls by the microarray software and increased separation between the location of the PCR primers and the microarray probes both led to reduced agreement. Microarray analysis suggested a floor effect in expression levels measured by RT-PCR for two genes. In conclusion, moderate mRNA expression levels with overlap in the location of PCR primers and microarray probes can yield good agreement between these two methods.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Etienne W,Meyer MH,Peppers J,Meyer RA Jrdoi
10.2144/04364ST02subject
Has Abstractpub_date
2004-04-01 00:00:00pages
618-20, 622, 624-6issue
4eissn
0736-6205issn
1940-9818journal_volume
36pub_type
杂志文章相关文献
BIOTECHNIQUES文献大全abstract：:We report a modified method of screening for point mutations using a PCR approach based upon the sensitivity of PCR to the 3' terminus of the primer. This method provides a sensitive screen when using either plasmid DNA or bacterial cell lysates. We have optimized the technique for general use to allow rapid screening...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1992-01-01 00:00:00
abstract：:We have developed a rapid, sensitive, and quantitative 96-well microplate-based nonradioactive immunoprecipitation/kinase assay to evaluate mitogen-activated protein kinase (MAPK) activity. Three quantitative nonradioactive imunoprecipitation/kinase assays of MAPK were demonstrated on a 96-well microplate coated with ...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/02322pf02
更新日期：2002-02-01 00:00:00
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
更新日期：2009-09-01 00:00:00
abstract：:Transfection of suspension cells has proven to be very difficult using conventional methods. Here, we present a simple and time-saving new transfection protocol wherein cell culture plates coated with chicken egg white are seeded with suspension cells prior to transfection. Our results demonstrate that coupling egg wh...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/000113914
更新日期：2012-08-01 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
pub_type： 杂志文章
doi：
更新日期：1988-09-01 00:00:00
abstract：:A method allowing the evaluation of the structure and the calculation of the volume of a biofilm, using an optical microscope, is proposed based on the linear relation between the intensity of a pixel in biofilm images grabbed on the x-y plane and the corresponding number of cells in the z direction, which allows the ...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/000112403
更新日期：2007-05-01 00:00:00
abstract：:A fully automated nucleic acid analysis system is described, which offers positive sample identification, improved sensitivity and reduced user interaction compared to conventional techniques. The system relies on the sequence-specific capture of DNA onto solid-phase particles, confirming product identity without the ...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/99271pf01
更新日期：1999-07-01 00:00:00
abstract：:Duracryl is a mechanically strong and elastic acrylamide-based matrix, useful for a wide variety of electrophoretic applications. The matrix is stable as a refrigerated solution for one year. Upon addition of appropriate catalysts, Duracryl forms a polymer-reinforced polyacrylamide gel matrix suitable for electrophore...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1992-04-01 00:00:00
abstract：:Presepsin is a 13-kDa N-terminal glycoprotein of CD14. Previously, agitation-induced increases in presepsin levels have been reported; however, the mechanism remains poorly understood. In this study, we aimed to reveal the mechanism of presepsin increase. The agitated plasma or serum was separated using gel exclusion ...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/btn-2020-0136
更新日期：2021-01-29 00:00:00
abstract：:An assay is described in which an oligonucleotide probe is specifically digested by lambda exonuclease only when it is annealed to its complementary sequence. In this assay, a cycling effect occurs whereby a small amount of target sequence acts as a specific co-factor in the enzymatic degradation of a larger number of...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1992-12-01 00:00:00
abstract：:DNA damage in the form of abasic sites, chemically altered nucleotides, and strand fragmentation is the foremost limitation in obtaining genetic information from many ancient samples. Upon cell death, DNA continues to endure various chemical attacks such as hydrolysis and oxidation, but repair pathways found in vivo n...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/000114307
更新日期：2015-07-01 00:00:00
abstract：:Gene expression studies require analysis of RNA, but isolation of total RNA from very small samples by traditional methods can be difficult and inefficient. The Absolutely RNA microprep kit provides a convenient method for isolating total RNA from small numbers of cells such as those harvested by laser capture microdi...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/01306pf01
更新日期：2001-06-01 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
pub_type： 杂志文章
doi：
更新日期：1991-01-01 00:00:00
abstract：:Specific amplification of nucleic acid sequences by PCR has been extensively used for the detection of gene rearrangements and gene expression. Although successful amplification of DNA sequences has been carried out with DNA prepared from formalin-fixed, paraffin-embedded (FFPE) tissues, there are only a few reports r...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1993-03-01 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
pub_type：
doi：
更新日期：1990-11-01 00:00:00
abstract：:Microarray platforms require analytical pipelines with modules for data pre-processing including data normalization, statistical analysis for identification of differentially expressed genes, cluster analysis, and functional annotation. We previously developed the Automated Microarray Data Analysis (AMDA, version 2.3....
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/0000113889
更新日期：2012-07-01 00:00:00
abstract：:In the design of oligonucleotide sequences for targeting DNA or RNA sequences, it can be difficult to identify sequences that will hybridize only to the intended target. The term "sequence-specific" or "sequence-nonspecific" is often used to describe the interactions of an oligonucleotide with a mixture of DNA or RNA....
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/96206pf01
更新日期：1996-06-01 00:00:00
abstract：:We describe a method for rapid radioactive labeling of PCR product. The method, employing the Klenow fragment of DNA polymerase I, consumes little product, requires no product purification and takes under 30 minutes. ...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1991-08-01 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
pub_type： 杂志文章
doi：
更新日期：1995-12-01 00:00:00
abstract：:Complete brain fixation can be achieved with transthoracic cardiac infusion without thoracotomy. Light and electron microscopy tissue sections reveal preservation of cytoplasmic and nuclear structure at all magnification levels. Punched samples were obtained from the fixed tissue specimens in precisely localized areas...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/000112003
更新日期：2005-10-01 00:00:00
abstract：:Advances in in vitro display and protein engineering yield therapeutics with affinities in the picomolar range. The Gyrolab® microfluidics platform uses the kinetic exclusion assay principle to measure subnanomolar solution affinities. This work describes application of the Gyrolab solution affinity module and the new...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/btn-2020-0080
更新日期：2020-09-01 00:00:00
abstract：:The concentration of proteins in cells is an important parameter that determines how a protein will interact with other proteins or pharmacological agents. Recent developments in Western blotting techniques have now made this a method of choice to measure protein concentration in complex solutions such as total cell e...
journal_title：BioTechniques
pub_type：
doi：
更新日期：1999-03-01 00:00:00
abstract：:The polymorphism of the fourth component of human serum complement (C4) is well established at the proteinic level; at the DNA level in the analysis of C4A and C4B gene polymorphism, the PCR technique is not widely and routinely used because it is time consuming and still presents reproducibility problems. This is a s...
journal_title：BioTechniques
pub_type：
doi：10.2144/01305st02
更新日期：2001-05-01 00:00:00
abstract：:Current protocols for screening proliferative factors for β cells ex vivo are time-consuming, require cell lines or dissociated islets, and often entail expensive specialized screening equipment. Here we present an efficient and lower cost alternative that utilizes intact mouse islets for the initial screening of prol...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/000114115
更新日期：2013-12-01 00:00:00
abstract：:3D printed biomaterials are increasingly used in cell cultures and drug screens. Given the ease of creating artificial tissues, will this technique revolutionize biomedicine and organ implants in the future? ...
journal_title：BioTechniques
pub_type： 新闻
doi：10.2144/btn-2017-0113
更新日期：2018-03-01 00:00:00
abstract：:We propose two methods for characterizing the spatio-temporal behavior of cell populations in culture. The first method, image auto-correlation microscopy (IACM), allows us to characterize the variation in the number of objects as a function of time, thus enabling the quantification of the clustering properties of cel...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/000112478
更新日期：2007-07-01 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
pub_type： 杂志文章
doi：10.2144/00295rr03
更新日期：2000-11-01 00:00:00
abstract：:The high-order structure of human chromosomes is an important biological question that is still under investigation. Studies have been done on imaging human mitotic chromosomes using mostly 2-D microscopy methods. To image micron-sized human chromosomes in 3-D, we developed a procedure for preparing samples for serial...
journal_title：BioTechniques
pub_type： 杂志文章
doi：10.2144/000114236
更新日期：2014-12-01 00:00:00
abstract：:Tandemly polymerized regulatory elements, antisense RNA segments or ribozymes are potentially useful in selective gene silencing. However, existing methods of tandemly polymerizing short DNA segments are laborious. We present a procedure that can create cloned arrays of 40-70 monomer units in two steps. We have create...
journal_title：BioTechniques
pub_type： 杂志文章
doi：
更新日期：1992-11-01 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
pub_type： 杂志文章
doi：10.2144/03351rr01
更新日期：2003-07-01 00:00:00
