Abstract：
:Analysis of single nucleotide polymorphisms by PCR with fluorescence resonance energy transfer (FRET) probes often can produce a result where the melting peak corresponding to perfectly matched sequence (A allele) has a smaller area than the peak corresponding to the allele with a mismatch (B allele). This imbalance can make it difficult to distinguish heterozygous individuals from BB homozygotes. These results suggested that the higher strength in the binding of the perfect match probe to the A allele could cause the selective amplification of the B allele, possibly by interfering with the elongation of the PCR product. In order to optimize the detection of heterozygotes in allelic discrimination assays with FRET probes, we tested several modifications aimed at minimizing the apparent interference of the probes with the amplification process. We observed, in agreement with our hypothesis, that lowering the probe concentration or adding the probes after the amplification step more accurately resolved heterozygotes.
journal_name
Mol Cell Probesjournal_title
Molecular and cellular probesauthors
Martínez-García A,Sastre I,Tenorio R,Bullido MJdoi
10.1016/j.mcp.2004.03.003subject
Has Abstractpub_date
2004-08-01 00:00:00pages
211-4issue
4eissn
0890-8508issn
1096-1194pii
S0890850804000325journal_volume
18pub_type
杂志文章abstract：BACKGROUND:Osteoarthritis (OA) is a frequent and incurable joint disease, inducing significant pain and seriously threatening to human health. It has been reported that microRNAs (miRNAs) play crucial roles on cancers and inflammatory diseases via cooperating with genes. However, the effect of miR-374a-3p/Wingless-type...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2020.101541
更新日期：2020-06-01 00:00:00
abstract：:The aim of the present study was to evaluate a mixed phase DNA hybridization assay for detection of Bordetella pertussis and Bordetella parapertussis in nasopharyngeal aspirates from patients with suspected pertussis. Among 179 consecutive patients with own or parental suspicion of pertussis, the diagnosis was confirm...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/0890-8508(90)90021-q
更新日期：1990-08-01 00:00:00
abstract：:Technologies that permit rapid investigation of DNA sequences, such as those containing single nucleotide polymorphisms (SNPs), are of great consequence to many sectors that perform molecular diagnostic analyses. We have developed a novel fluorescent oligonucleotide probe technology, termed HyBeacons, which provides a...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1006/mcpr.2001.0384
更新日期：2001-12-01 00:00:00
abstract：:The ability to preserve and transport human cells in a stable medium over long distances is critical to collaborative efforts and the advancement of knowledge in the study of human disease. This is particularly important in the study of rare diseases. Recently, advancements in the understanding of renal ciliopathies h...
journal_title：Molecular and cellular probes
pub_type： 杂志文章,评审
doi：10.1016/j.mcp.2021.101694
更新日期：2021-01-08 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
abstract：:We identified multiple paternity in 52.9% of the clutches of Hermann's tortoise Testudo hermanni boettgeri using polymorphic microsatellite markers. In addition we demonstrated sperm storage across seasons. DNA was extracted from the amniotic fluid adhering to the eggshell's inner surface, a procedure suitable for eas...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2015.05.009
更新日期：2015-08-01 00:00:00
abstract：:Advanced reflectance-based optical techniques for in vivo imaging often suffer from low contrast between neoplastic and normal tissue and are unable to image early biomolecular changes associated with carcinogenesis, thus limiting their clinical value. In this study, we exploit the resonance light scattering property ...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2007.06.010
更新日期：2008-02-01 00:00:00
abstract：:Serum lipid profile which is determined by genotype-phenotype relationship plays a significant role in the development of cardiovascular disease. Upstream stimulatory factor 1 (USF1), has been reported to be associated with serum lipid levels in different population, hence, this study investigated the association of v...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2020.101687
更新日期：2020-12-08 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：:Double polymerase chain reaction (PCR) assays with nested primers have been applied in a routine laboratory for the diagnosis of herpes-, pesti- and retroviral infections of animals. Various methods and tools have been tested to prevent and to eliminate false positive results as well as to visualize the PCR products (...
journal_title：Molecular and cellular probes
pub_type： 杂志文章,评审
doi：10.1006/mcpr.1993.1035
更新日期：1993-06-01 00:00:00
abstract：:The G-->A transition at nucleotide 21881 of the human catechol-O-methyltransferase (COMT) gene represents a functional genetic polymorphism (Val158Met), rendering an enzyme with reduced activity that has been associated with psychiatric disorders and estrogen-related cancers. A new method for the detection of this pol...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2006.12.001
更新日期：2007-06-01 00:00:00
abstract：:We have applied the temperature-dependent single-stranded conformational polymorphism (SSCP) analysis for characterization of influenza A virus cDNA fragments. A series of primers were synthesized on the basis of the comparison of hemagglutinin and PB2 gene sequences of different origin. RT-PCR reactions were run usin...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2008.09.003
更新日期：2008-10-01 00:00:00
abstract：:A random amplified polymorphic DNA (RAPD) technique using 16 decamer oligonucleotide primers was employed to characterize isolates of Schistosoma japonicum from seven geographical locations (Sj1: Zhejiang; Sj2: Anhui; Sj3: Jiangxi; Sj4: Hunan; Sj5: Hubei; Sj6: Sichuan; Sj7: Yunnan) of the People's Republic of China. D...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1006/mcpr.1996.0048
更新日期：1996-10-01 00:00:00
abstract：:Quantum dot (QD) molecular beacons were explored for sequence-specific DNA detection. The effectiveness of multiple linkage strategies and fluorescence quenchers were compared in hybridization-based assays. To compare linkage strategies, covalent amide linkage and streptavidin-biotin binding were used to link semicond...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2006.09.001
更新日期：2007-04-01 00:00:00
abstract：:Holstein haplotype (HH) 1, 3 and 4 are lethal mutations, responsible for early embryonic losses in Holstein Friesian (HF) cattle, worldwide. Three PCR based assays - tetra Amplification Refractory Mutation System PCR, PCR primer induced restriction analysis and PCR-restriction fragment length polymorphism techniques f...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2019.101503
更新日期：2020-04-01 00:00:00
abstract：:The development of a rapid, specific, and sensitive SYBR Green I-based duplex real-time quantitative PCR assay is described for the simultaneous detection of porcine epidemic diarrhea virus (PEDV) and porcine circovirus type 3 (PCV3). The assay specifically detected PEDV and PCV3, with no fluorescence detected for oth...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2019.02.002
更新日期：2019-04-01 00:00:00
abstract：:Hypertriglyceridemia (HTG) plays an important role in the development and progression of atherosclerosis. It is inherited in an autosomal dominant pattern with a frequency of approximately 1:1,000,000 worldwide. Previous study has demonstrated that more than six genes underlie this disorder. In addition, copy number v...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2017.11.002
更新日期：2018-02-01 00:00:00
abstract：:A simple and rapid method, PCR-restriction enzyme analysis (PCR-RE) for BK virus (BKV) typing was developed, based on the presence of type-specific restriction enzyme sites in a 327 bp PCR-generated fragment which partially encodes the VP1 protein. The enzymes, Alu I, Xmn I and Ava II, were used to digest the PCR prod...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1006/mcpr.1993.1047
更新日期：1993-08-01 00:00:00
abstract：:The ability to rapidly and reliably genotype mice is an important concern. Traditional methods employ labour intensive and time consuming techniques such as test crossing, gel electrophoresis or nucleic acid hybridization. Here we show that a new molecular biology workstation, the WAVE DNA Fragment Analysis System, ca...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1006/mcpr.1999.0239
更新日期：1999-06-01 00:00:00
abstract：:Although enteroviral infections occur frequently during childhood, the circulation of particular serotypes has never been studied in Greece. The objectives of the present report were molecular detection and identification of human enteroviruses in children admitted with nonspecific febrile illness or meningitis to a u...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2011.06.001
更新日期：2011-10-01 00:00:00
abstract：:TTR amyloidosis (ATTR) is a fatal condition caused by extracellular deposits of misfolded transthyretin. Patients often present with cardiac disease, but manifestations may also involve other organs including the peripheral nervous system. ATTR is considered familial when heterozygous mutations in the TTR gene are pre...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2018.08.005
更新日期：2018-10-01 00:00:00
abstract：:Non-pathogenic Burkholderia thailandensis may be used as a model for Burkholderia pseudomallei due to the genetic similarity of these species. Moreover, the experimental manipulation of B. thailandensis is safer. In this study, we constructed recombinant flagellin protein fragments of B. thailandensis E264 (FLAG300, F...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2012.11.001
更新日期：2013-04-01 00:00:00
abstract：:The progressive myoclonus epilepsy of Lafora type (LD) is an autosomal recessive disorder caused by mutations in the EPM2A gene. We demonstrated recently that EPM2A encodes a dual-specificity phosphatase that is primarily associated with polyribosomes. In the present study, we screened for mutations in the EPM2A gene ...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1006/mcpr.2001.0371
更新日期：2001-10-01 00:00:00
abstract：:Mycoplasma infections are of great concern in avian medicine, because they cause economic losses in commercial poultry production. A multiplex polymerase chain reaction (PCR) was optimized to simultaneously detect four pathogenic species of avian mycoplasmas. Four sets of oligonucleotide primers specific for Mycoplasm...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1006/mcpr.1997.0108
更新日期：1997-06-01 00:00:00
abstract：:Porcine epidemic diarrhea (PED) caused by porcine epidemic diarrhea virus (PEDV) is a highly contagious intestinal disease, resulting in substantial economic losses to the swine industry worldwide. In this study, three assays, namely a conventional reverse transcription-polymerase chain reaction (RT-PCR), a SYBR Green...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2017.02.002
更新日期：2017-06-01 00:00:00
abstract：:Strand displacement amplification (SDA) is an isothermal, in vitro method of amplifying a target DNA sequence. We performed SDA in the presence of a 5'-32P-oligodeoxynucleotide detector probe that contains a target binding sequence at its 3'-end and a recognition site for the restriction enzyme HincII at its 5'-end wh...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1006/mcpr.1995.0062
更新日期：1995-12-01 00:00:00
abstract：:DNA samples of 2303 individuals from nine different population groups were screened for variant -175g-->t in the promoter region of the low-density lipoprotein receptor (LDLR) gene. The -175g-->t variant detected at carrier frequencies of 3-10% in different African population groups was absent in the Caucasian and Asi...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/s0890-8508(03)00050-1
更新日期：2003-08-01 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：:A DNA amplification procedure using heat stable Taq polymerase and the polymerase chain reaction is described for the detection of Pseudomonas aeruginosa in specimens from cystic fibrosis patients. A set of primers was selected on the basis of the nucleotide sequence of the algD gene encoding GDP mannose dehydrogenase...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/0890-8508(92)90005-i
更新日期：1992-08-01 00:00:00
abstract：BACKGROUND:Stroke is a leading cause of mortality worldwide. Rac-MAPK kinase 6 (Map2k6) plays important roles in cell proliferation and apoptosis. However, the role played by Map2k6 in stroke injury and the underlying mechanism of action remain unknown. METHODS:Mice received cerebral ischemia/reperfusion (I/R) injurie...
journal_title：Molecular and cellular probes
pub_type： 杂志文章
doi：10.1016/j.mcp.2019.101478
更新日期：2020-02-01 00:00:00
