Abstract:
:We describe a rapid, automated method for direct detection of known single-base changes in genomic DNA. Fluorescence-based DNA minisequence analysis is employed in a template-dependent reaction which involves a single nucleotide extension of an oligonucleotide primer by the correct fluorescently-tagged dideoxynucleotide chain terminator. Detection following electrophoresis on denaturing acrylamide gels is facilitated by alkaline phosphatase treatment of reaction products after extension followed by isopropanol precipitation of the dye-tagged, single-base-extended primer to remove unincorporated deoxynucleotides. Fluorescence analysis of the incorporated dye tag reveals the identity of the template nucleotide immediately 3' to the primer site. This technique does not require radioactivity or biotinylated PCR product, relies on the incorporation of a single dideoxynucleotide terminator to extend the primer by one nucleotide and takes advantage of the sensitivity of fluorescent terminators developed for automated DNA sequence analysis. As a demonstration, we have applied the assay to human genomic DNA for detection of the sickle mutation in the beta-globin gene, and have also examined feasibility for simultaneous delineation using a multiplex-like strategy in a single gel-lane of some of the most common beta-thalassemia mutations in the Mediterranean basin.
journal_name
Mol Cell Probesjournal_title
Molecular and cellular probesauthors
Kobayashi M,Rappaport E,Blasband A,Semeraro A,Sartore M,Surrey S,Fortina Pdoi
10.1006/mcpr.1995.0027subject
Has Abstractpub_date
1995-06-01 00:00:00pages
175-82issue
3eissn
0890-8508issn
1096-1194pii
S0890-8508(85)70027-8journal_volume
9pub_type
杂志文章abstract::Non-small-cell lung carcinoma (NSCLC) accounts for approximately 80% of lung cancers with a high metastatic potential. Elucidating the mechanism of NSCLC metastasis will provide new promising targets for NSCLC therapy and benefit its prognosis. Plasmacytoma variant translocation 1 (PVT1) has been proven to be overexpr...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2020.101652
更新日期:2020-12-01 00:00:00
abstract::To explore the genetic divergence and phylogeny of Chinese indigenous sheep breeds, in the current study, we analyzed the polymorphisms of 5 structural loci in ten sheep populations, including Sishui Fur, Sunite, Wurank, Bayinbuluke, Altay, Small-Tailed Han, Wadi, Tan, Tong and Hu sheep. The data were then compared wi...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2017.11.006
更新日期:2018-02-01 00:00:00
abstract::Metastasis and recurrence of breast cancer remain significant clinical problems. The expression level of CD44 protein is higher in breast cancer-initiating cancer stem cells; therefore, the early detection of CD44 using a sensitive diagnostic probe is important for breast cancer diagnosis and therapeutic purposes. In ...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2015.05.014
更新日期:2015-12-01 00:00:00
abstract::A full-length complementary DNA (cDNA; designated Hc-stp-1) encoding a serine/threonine phosphatase (Hc-STP-1) was isolated from Haemonchus contortus, a strongylid nematode parasite of small ruminants. Hc-stp-1 was shown to be transcribed in males of both adults and fourth-stage larvae, but not in females, early larva...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2010.02.001
更新日期:2010-08-01 00:00:00
abstract::Human metapneumovirus (hMPV) is a prevalent pathogen worldwide and causes various respiratory infections. Although it is a critical pathogen in pediatric patients, it is unclear how it enters host cells. In this study, we focused on hMPV cell entry using two kinds of cell lines (Vero E6 and LLC-MK2), which are most co...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2016.06.003
更新日期:2016-08-01 00:00:00
abstract:BACKGROUND:Androgen receptor (AR) and long non-coding RNAs (lncRNA) play important roles in the initiation and progression of prostate cancer (PCa). The present study was designed to investigate whether lncRNA growth arrest-specific 5 (GAS5) is involved in the regulation of dexamethasone on the proliferation of AR+ PCa...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2020.101607
更新日期:2020-10-01 00:00:00
abstract::For the detection of Chlamydia pneumoniae by polymerase chain reaction (PCR) in respiratory samples, an internal control was constructed to monitor the efficiency of amplification in each reaction. The internal control was designed in a way that the same primer pair can be used to amplify the internal control and targ...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1006/mcpr.1998.0180
更新日期:1998-08-01 00:00:00
abstract::Infection of the cervix uteri with various types of human papillomaviruses is generally considered a necessary factor in the etiology of cancer of the cervix uteri. In many human populations throughout the world, approximately 90% of cervical carcinomas are found to harbour HPV genomes, as judged by Southern blot hybr...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/0890-8508(90)90013-p
更新日期:1990-04-01 00:00:00
abstract::The real-time PCR-HRM analysis was developed for the detection and discrimination of the quarantine nematode Bursaphelenchus xylophilus and Bursaphelenchus mucronatus. A set of primers was designed to target the ITS region of rDNA. The results have demonstrated that this analysis is a valuable tool for differentiation...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2016.02.003
更新日期:2016-04-01 00:00:00
abstract::DNA probes were used to examine tetracycline-resistant Gram-negative bacteria (281 strains representing eight species) from catfish ponds. The isolates, which did not previously hybridize with the Tet A, B and C determinants, were examined for the presence of tetracycline-resistance Tet D and Tet E determinants. The d...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/s0890-8508(95)91572-9
更新日期:1995-10-01 00:00:00
abstract::At some stages of development, it is impossible to identify the porcine nodular worms Oesophagostomum dentatum and O. quadrispinulatum to the species level using morphological parameters. A molecular approach utilizing genetic markers in the second internal transcribed spacer (ITS-2) of ribosomal (r) DNA was developed...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1006/mcpr.1997.0097
更新日期:1997-04-01 00:00:00
abstract::Using an acute promyelocytic leukemia (APL) preclinical model, we show that oncogene-specific PCR (Polymerase Chain Reaction)-based assays allow to evaluate the efficacy of immunotherapy combining all-trans retinoic acid (ATRA) and a DNA-based vaccine targeting the promyelocytic leukemia-retinoic acid receptor alpha (...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2012.08.001
更新日期:2013-02-01 00:00:00
abstract::The manual gravimetric drying moisture determination methods currently employed by most mineral processing plants fail to provide timely and accurate information required for automatic control. The costs associated with transporting and handling concentrates still represent a major portion of the overall treatment pri...
journal_title:Molecular and cellular probes
pub_type: 杂志文章,评审
doi:10.1006/mcpr.2002.0444
更新日期:2002-12-01 00:00:00
abstract::To find a specific method for HLA-B27 typing, we tested an HLA-B27-specific polymerase chain reaction. This method was used for screening 100 randomly selected blood donors, 10 of them being HLA-B27 positive. A flow cytometric method and this PCR method were compared. ...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1006/mcpr.1997.0112
更新日期:1997-08-01 00:00:00
abstract::Beet Necrotic Yellow Vein Virus (BNYVV) was detected by enzyme-linked immunosorbent assay (ELISA) and RNA/DNA dot hybridization using either radiolabelled or non-radioactive probes. Dot hybridization specifically distinguished isolates that could not be distinguished by ELISA. The detection thresholds for ELISA, hybri...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/0890-8508(89)90026-1
更新日期:1989-06-01 00:00:00
abstract:OBJECTIVES:As one of the most serious malignant carcinomas that threaten the life of sufferers constantly, gastric cancer has attracted a lot of interest among researchers. miR-34a, a member of hundreds of microRNAs (miRNAs), has been elucidated to exert a suppressive role in gastric cancer tumorgenesis based on previo...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2020.101601
更新日期:2020-10-01 00:00:00
abstract::Spinal muscular atrophy (SMA), the leading genetic cause of death in childhood, is an autosomal recessive neuromuscular disorder characterized by progressive muscle weakness, associated with deletions of the survival motor neuron 1 (SMN1) gene. Approximately 94% of SMA patients carry homologous deletions of SMN1 exon(...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2009.12.001
更新日期:2010-06-01 00:00:00
abstract::Several dyes are now known which intercalate into double-stranded DNA with fluorescence enhancement. One such is TOTO (1,1'-(4,4,7,7-tetramethyl-4,7-diazaundecamethylene)-bis-4-[3-meth yl-2,3- dihydro-(benzo-1,3-thiazole)-2-methlyidene]-quinolinium tetraiodide), a homodimer of thiazole orange, whose complexes with DNA...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1006/mcpr.1994.1035
更新日期:1994-06-01 00:00:00
abstract::A polymerase chain reaction (PCR) method specific for Mycoplasma gallisepticum (MG) was evaluated. The PCR method was found to detect as few as two colour changing units (CCU) of MG and did not give false positive reactions with other avian mycoplasmas. In chickens inoculated with either MG or Mycoplasma synoviae (MS)...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1006/mcpr.1993.1068
更新日期:1993-12-01 00:00:00
abstract::One pair of high-sensitive polymerase chain reaction (PCR) primers for Cryptosporidium parvum was constructed based on the sequence of random amplified polymorphic DNA. PCR with this primer pair amplified only the DNA of C. parvum, not the control DNA including Cryptosporidium muris. This primer pair had most advantag...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1006/mcpr.1999.0280
更新日期:2000-02-01 00:00:00
abstract::Several systems for isolating viruses from environmental samples have been tested. The most promising method is based on genomic amplification. The authors attempted to detect adenovirus in nucleic-acid extracts from the Seine River estuary by a two-step amplification of a 220-bp segment of the conserved coding region...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1006/mcpr.1998.0166
更新日期:1998-06-01 00:00:00
abstract::The first report of pertactin-negative variants of Bordetella pertussis in the United States has raised questions about the role of acellular pertussis vaccines in the recent increase of pertussis cases. Our laboratory utilized a sequence-based method to identify mutations in the pertactin gene responsible for these v...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2013.12.003
更新日期:2014-08-01 00:00:00
abstract::Previous studies have demonstrated that insulin-like growth factor-I (IGF-1) and reactive oxygen species (ROS) are involved in the development and progression of various cancers. However, their regulatory mechanism remains unknown. In this study, we treated cancer cells (HeLa, HepG2 and SW1116 cells) and normal cells ...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2020.101583
更新日期:2020-08-01 00:00:00
abstract::We have developed a new methodology for fluorescence turn-on detection of DNA methyltransferase (MTase) activity based on terminal deoxynucleotidyl transferase (TdT) using a thioflavin T probe. This method is highly selective and sensitive. The fluorescence intensity was direct proportion to Dam MTase concentration in...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2016.02.001
更新日期:2016-04-01 00:00:00
abstract::Natalizumab is a humanized monoclonal antibody against the alpha4 chain of the alpha4beta1 and alpha4beta7 integrin heterodimers used with high effectiveness in the treatment of multiple sclerosis. The use of this drug can unfortunately be associated with the onset of progressive multifocal leukoencephalopathy, a poss...
journal_title:Molecular and cellular probes
pub_type: 杂志文章,评审
doi:10.1016/j.mcp.2014.11.007
更新日期:2015-02-01 00:00:00
abstract::We have developed and evaluated an ELISA-based detection method for PCR-amplified HIV-1 DNA. The assay uses two oligonucleotide probes which are end-labelled at the 5'-end with biotin or digoxigenin, respectively. Upon solution hybridization of these probes which react with the same strand of amplified DNA product, th...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1006/mcpr.1993.1054
更新日期:1993-10-01 00:00:00
abstract::Diabetic cardiomyopathy (DCM) is a common complication of diabetes mellitus that can cause many severe symptoms, such as heart failure, arrhythmia, and sudden death. However, the molecular mechanisms underlying cardiac dysfunction in DCM remain elusive. In this study, we found that miR-410-5p was increased in the myoc...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2020.101558
更新日期:2020-08-01 00:00:00
abstract::Identification of sources Campylobacter infection in the poultry houses is in general problematic due to the lack of reliable methods to detect campylobacteria in environmental samples. Detection of campylobacteria in environmental samples by conventional culture methods is difficult and of limited sensitivity due to ...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1006/mcpr.2002.0434
更新日期:2002-10-01 00:00:00
abstract::The vast majority of first-trimester pregnancy losses are the consequence of numerical aberrations in fetal chromosomes, which may involve nearly all chromosomes. Although commercial probes for all chromosomes are available for multiplex ligation-dependent probe amplification (MLPA) and fluorescence in situ hybridizat...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1016/j.mcp.2019.101422
更新日期:2019-08-01 00:00:00
abstract::As part of the development of the ligase chain reaction (LCR) into a tool which can be used by a wide variety of researchers, we have investigated several analytical detection systems for the products of this amplification reaction. While early work with this technology has used gel electrophoresis to separate the LCR...
journal_title:Molecular and cellular probes
pub_type: 杂志文章
doi:10.1006/mcpr.1993.1027
更新日期:1993-06-01 00:00:00