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 expression of chloramphenicol acetyltransferase (CAT) on a single cell basis. The results show that myocardial cells take up and express transfected DNA approximately 7 times less efficiently than non-myocardial cells. The immunofluorescence analysis allows the biochemical level of CAT activity expressed under control of different transcriptional promoters to be compared on a cell type by cell type basis. Thus, the relative transcriptional strength of two promoters can be estimated for a specific cell type within a heterogeneous population of cells.
journal_name
Biotechniquesjournal_title
BioTechniquesauthors
Antin PB,Mar JH,Ordahl CPsubject
Has Abstractpub_date
1988-07-01 00:00:00pages
640-2, 645-9issue
7eissn
0736-6205issn
1940-9818journal_volume
6pub_type
杂志文章,评审相关文献
BIOTECHNIQUES文献大全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:
更新日期:1989-11-01 00:00:00
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
更新日期:2003-09-01 00:00:00
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
更新日期:2020-10-01 00:00:00
abstract::We describe a rapid and sensitive method for the detection of nucleotide sequence variation that can be used for large-scale screening of population markers. Denaturing gradient gel electrophoresis (DGGE) detects sequence variants of amplified fragments by the differences in their melting behavior. DGGE detects most s...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/99275rr02
更新日期:1999-11-01 00:00:00
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:
更新日期:1994-08-01 00:00:00
abstract::Quantitative PCR and reverse transcription PCR (RT-PCR) are widely used in biomedical, industrial and other research applications to determine the number of RNA or DNA molecules of a specific type and/or sequence in a sample of interest. We have developed an assay system to accurately quantitate PCR products that util...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1995-05-01 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
pub_type: 杂志文章
doi:10.2144/000114217
更新日期:2014-10-01 00:00:00
abstract::We describe a simple and efficient system for epitope mapping by cloning random gene fragments into a specially designed gIIIp-based phage display vector. DNA encoding the antigen of interest is PCR-amplified and partially digested with DNaseI to generate 50-150-bp-long fragments, which are polished with T4 DNA polyme...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/99272st04
更新日期:1999-08-01 00:00:00
abstract::SOP3 is a web-based software tool for designing oligonucleotide primers for use in the analysis of single nucleotide polymorphisms (SNPs). Accessible via the Internet, the application is optimized for developing the PCR and sequencing primers that are necessary for Pyrosequencing. The application accepts as input gene...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/05381RR01
更新日期:2005-01-01 00:00:00
abstract::Human multiple tissue Western (MTW) blots are premade immunoblots prepared using proteins isolated from adult human tissue. The proteins are isolated from whole tissue homogenates under conditions designed to minimize proteolysis and to ensure maximal representation of tissue-specific proteins. Sodium dodecyl sulfate ...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1994-11-01 00:00:00
abstract::Tissue engineering scaffolds have emerged as a powerful tool within regenerative medicine. These materials are being designed to create environments that promote regeneration through a combination of: (i) scaffold architecture, (ii) the use of scaffolds as vehicles for transplanting progenitor cells, and/or (iii) loca...
journal_title:BioTechniques
pub_type: 杂志文章,评审
doi:10.2144/000113754
更新日期:2011-10-01 00:00:00
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:
更新日期:1994-04-01 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::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
更新日期:2014-07-01 00:00:00
abstract::Selective delivery of genes to ocular tissues in vivo has been a long sought after goal for potential gene therapy of ocular disease. The gene gun was considered for this purpose because of its ability to focally transfer DNA to cells through gold microparticles coated with DNA. Through experimentation, we optimized a...
journal_title:BioTechniques
pub_type:
doi:10.2144/97233st06
更新日期:1997-09-01 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
pub_type: 杂志文章
doi:10.2144/000114330
更新日期:2015-09-01 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
pub_type: 杂志文章
doi:10.2144/btn-2019-0148
更新日期:2020-06-01 00:00:00
abstract::A method for conserving primers and differently labeled fluorogenic hydrolysis (i.e., TaqMan) probes at ambient conditions is presented. Primers and hydrolysis probes with four different fluorophore-quencher combinations (6- FAM-BHQ1, HEX-BHQ1, ROX-BHQ650, and Cy5-BHQ2) were mixed with trehalose and xanthan at final c...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000114207
更新日期:2014-09-01 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
pub_type: 杂志文章
doi:
更新日期:1988-11-01 00:00:00
abstract::Microarray expression analysis has become one of the most widely used functional genomics tools. Efficient application of this technique requires the development of robust and reproducible protocols. We have optimized all aspects of the process, including PCR amplification of target cDNA clones, microarray printing, p...
journal_title:BioTechniques
pub_type: 杂志文章,评审
doi:10.2144/00293bi01
更新日期:2000-09-01 00:00:00
abstract::Autoantibodies directed against the 68-kDa (U1) ribonucleoprotein antigen are mainly found in sera of patients with mixed connective tissue disease. The corresponding cDNA was fragmented into four regions coding for the major antigenic epitopes A', B', C' and D'. All the epitopes were subcloned and expressed as fusion...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1991-09-01 00:00:00
abstract::The relative spatial distribution of cells in a solid tumor contributes to development of malignancy, yet the details of this process remain poorly understood. To elucidate these mechanisms, the ability to extract and analyze the entire DNA content of individual cells whose precise location in the tumor is known is re...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000113860
更新日期:2012-04-01 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
pub_type: 杂志文章
doi:10.2144/000112724
更新日期:2008-04-01 00:00:00
abstract::For a feasible and cost-effective impedance measurement of cellular alterations in real-time, we combined commercially available microelectrode arrays (MEAs), consisting of 60 microelectrodes, with a conventional impedance analyzer. For proof of principle, a breast carcinoma cell line (MCF-7) was cultured on MEAs, and...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000112254
更新日期:2006-10-01 00:00:00
abstract::Microsatellite sequences are important markers for population genetics studies. In the past, the development of adequate microsatellite primers has been cumbersome. However with the advent of next-generation sequencing technologies, marker identification in genomes of non-model species has been greatly simplified. Her...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/000114104
更新日期:2013-11-01 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::Metabolic and electrical coupling through gap junction channels is implicated in cell differentiation, tissue homeostasis, and electrotonic propagation of signals in excitable tissues. The characterization of gating properties of these channels requires electrophysiological recordings at both single- and multiple-chan...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/03345rr03
更新日期:2003-05-01 00:00:00
abstract::Hematopoietic stem cells (HSCs) express Mdr1a/1b and Bcrp1/Abcg2, which are members of the ATP binding cassette transporter family. Mice lacking both Mdr1-type genes (Mdr1a and Mdr1b) or Bcrp1 had normal hematopoietic development, but it has been unclear whether Mdr1a/1b and Bcrp1 play redundant roles in hematopoiesis...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/03356ss04
更新日期:2003-12-01 00:00:00
abstract::S-100B is used in melanoma follow-up. This serum biomarker is also present in adipocytes; therefore, subcutaneous adipocytes trapped in the needle before performing a venipuncture could contaminate the serum. The aim was to study the influence of adipocyte contamination on blood samples used for S-100B analysis, possi...
journal_title:BioTechniques
pub_type: 杂志文章
doi:10.2144/btn-2019-0147
更新日期:2020-11-01 00:00:00
abstract::We have developed a procedure for undertaking a Microtox-based test by coupling microplate and microluminometric technologies. Sample dilutions are prepared in a 96-well polystyrene microplate kept at 15 degrees C, while the Microtox reagent and diluent are placed in an opaque, microluminometry-compatible 96-well micr...
journal_title:BioTechniques
pub_type: 杂志文章
doi:
更新日期:1994-05-01 00:00:00