Genetic analysis, techniques and applications最新文献_第10页

The application of microwave denaturation in comparative genomic hybridization 微波变性在比较基因组杂交中的应用

Genetic analysis, techniques and applications Pub Date : 1996-11-01 DOI: 10.1016/S1050-3862(96)00162-3

Maurice de Meulemeester , Agnes Vinka , Marja Jakobs , Mario Hermsen , Marja Steenman , Rosalyn Slates, Axel Dietrich , Marcel Mannensa

{"title":"The application of microwave denaturation in comparative genomic hybridization","authors":"Maurice de Meulemeester , Agnes Vinka , Marja Jakobs , Mario Hermsen , Marja Steenman , Rosalyn Slates, Axel Dietrich , Marcel Mannensa","doi":"10.1016/S1050-3862(96)00162-3","DOIUrl":"10.1016/S1050-3862(96)00162-3","url":null,"abstract":"<div><p>Comparative genomic hybridization (CGH) is a powerful tool for analyzing unbalanced chromosomal rearrangements in a variety of tissues. However, the reproducibility of the technique is poor. We have developed an alternative protocol involving microwave denaturation of the metaphase chromosome preparations prior to the hybridization step. The advantage of this method for CGH is the retention of the morphology of the chromosomes and hence an improved chromosome banding pattern. Furthermore, it results in a cansistently strong hybridization which is not dependent on the batch of lymphocytes used to obtain the metaphase chromosome spreads. This procedure has also proved to be applicable to nucleic acid hybridizations in general. The protocol, its application and the results of this method in CGH is discussed. Furthermore preliminary results of this method in paint and DNA probe hybridizations to chromosome spreads and to RNA in tissue sections are presented.</p></div>","PeriodicalId":77142,"journal":{"name":"Genetic analysis, techniques and applications","volume":"13 5","pages":"Pages 129-133"},"PeriodicalIF":0.0,"publicationDate":"1996-11-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S1050-3862(96)00162-3","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19980253","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 26

In situ detection of tandem DNA repeat length 串联DNA重复长度的原位检测

Genetic analysis, techniques and applications Pub Date : 1996-11-01 DOI: 10.1016/S1050-3862(96)00155-6

Ron Yaar, Przemyslaw Szafranski, Charles R. Cantor, Cassandra L. Smith

引用次数: 4

Long range-inverse PCR (LR-IPCR): extending the useful range of inverse PCR Long range-inverse PCR (LR-IPCR):扩大了逆转录PCR的应用范围

Genetic analysis, techniques and applications Pub Date : 1996-11-01 DOI: 10.1016/S1050-3862(96)00161-1

Bernhard F. Benkel, Ying Fong

引用次数: 37

Laser desorption mass spectrometry for point mutation detection 激光解吸质谱法用于点突变检测

Genetic analysis, techniques and applications Pub Date : 1996-10-01 DOI: 10.1016/S1050-3862(95)00154-9

N.I Taranenko , K.J Matteson , C.N Chung , Y.F Zhu , L.Y Chang , S.L Allman , L Haff , S.A Martin , C.H Chen

{"title":"Laser desorption mass spectrometry for point mutation detection","authors":"N.I Taranenko , K.J Matteson , C.N Chung , Y.F Zhu , L.Y Chang , S.L Allman , L Haff , S.A Martin , C.H Chen","doi":"10.1016/S1050-3862(95)00154-9","DOIUrl":"10.1016/S1050-3862(95)00154-9","url":null,"abstract":"<div><p>A point mutation can be associated with the pathogenesis of inherited or acquired diseases. Laser desorption mass spectrometry coupled with allele specific polymerase chain reaction (PCR) was first used for point mutation detection. G551D is one of several mutations of the cystic fibrosis transmembrane conductance regulator (CFTR) gene present in 1–3% of the mutant CFTR alleles in most European populations. In this work, two different approaches were pursued to detect G551D point mutation in the cystic fibrosis gene. The strategy is to amplify the desired region of DNA template by PCR using two primers that overlap one base at the site of the point mutation and which vary in size. If the two primers based on the normal sequence match the target DNA sequence, a normal PCR product will be produced. However, if the alternately sized primers that match the mutant sequence recognize the target DNA, an abnormal PCR product will be produced. Thus, the mass spectrometer can be used to identify patients that are homozygous normal, heterozygous for a mutation or homozygous abnormal at a mutation site. Another approach to identify similar mutations is the use of sequence specific restriction enzymes which respond to changes in the DNA sequence. Mass spectrometry is used to detect the length of the restriction fragments generated by digestion of a PCR generated target fragment.</p></div>","PeriodicalId":77142,"journal":{"name":"Genetic analysis, techniques and applications","volume":"13 4","pages":"Pages 87-94"},"PeriodicalIF":0.0,"publicationDate":"1996-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S1050-3862(95)00154-9","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19913721","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 28

A biotinylated MutS fusion protein and its use in a rapid mutation screening technique 一种生物素化MutS融合蛋白及其在快速突变筛选技术中的应用

Genetic analysis, techniques and applications Pub Date : 1996-10-01 DOI: 10.1016/S1050-3862(95)00160-3

Daniel H. Geschwind , Richard Rhee , Stanley F. Nelson

引用次数: 19

Identification and mapping of a putative bombesin receptor gene on human chromosome 17q21.3+ 人染色体17q21.3+上一个推定的炸弹素受体基因的鉴定和定位

Genetic analysis, techniques and applications Pub Date : 1996-10-01 DOI: 10.1016/S1050-3862(95)00158-1

Uddhav Kelavkar , Ken Abel , Diane Miller , James Murtagh , Ketan Shah

引用次数: 0

Application of repetitive sequence-based PCR (inter-LINE PCR) for the analysis of genomic rearrangements and for the genome characterization on different taxonomic levels 基于重复序列的PCR (inter-LINE PCR)在基因组重排分析和不同分类水平上的基因组表征中的应用

Genetic analysis, techniques and applications Pub Date : 1996-10-01 DOI: 10.1016/S1050-3862(95)00157-3

Jan Smida , Stefan Leibhard , Anja M. Nickel , Friederike Eckardt-Schupp , Ludwig Hieber

引用次数: 15

Detection of hepatitis B virus DNA in serum samples via nested PCR and MALDI-TOF mass spectrometry 巢式PCR和MALDI-TOF质谱法检测血清乙型肝炎病毒DNA

Genetic analysis, techniques and applications Pub Date : 1996-09-01 DOI: 10.1016/1050-3862(95)00151-4

C. Jurinke , B. Zöllner , H.-H. Feucht , A. Jacob , J. Kirchhübel , A. Lüchow , D. van den Boom , R. Laufs , H. Köster

{"title":"Detection of hepatitis B virus DNA in serum samples via nested PCR and MALDI-TOF mass spectrometry","authors":"C. Jurinke , B. Zöllner , H.-H. Feucht , A. Jacob , J. Kirchhübel , A. Lüchow , D. van den Boom , R. Laufs , H. Köster","doi":"10.1016/1050-3862(95)00151-4","DOIUrl":"10.1016/1050-3862(95)00151-4","url":null,"abstract":"<div><p>In a blind study, nested polymerase chain reaction (PCR) was performed with control DNA and DNA preparations from serum samples of six patients. The detection limit was determined to be 100 molecules of template in 1 ml of serum. Hepatitis B virus (HBV) related products of nested PCR were purified by ultrafiltration and immobilisation on streptavidin coated magnetic beads. The immobilized PCR products were denatured from the beads and analyzed via matrix assisted laser desorption/ionisation time-of-flight (MALDI-TOF) mass spectrometry. The results of MALDI-TOF MS analysis were in agreement with the results obtained by polyacrylamide gel electrophoresis (PAGE) and with the data obtained by serological analysis. The detection strategy introduced here has a high potential for automation and represents a fast and reliable method of detection for HBV DNA in serum without the need for time consuming gel electrophoresis and labeling or hybridization procedures.</p></div>","PeriodicalId":77142,"journal":{"name":"Genetic analysis, techniques and applications","volume":"13 3","pages":"Pages 67-71"},"PeriodicalIF":0.0,"publicationDate":"1996-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/1050-3862(95)00151-4","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19896085","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 26

HLA allele selection for designing peptide vaccines HLA等位基因选择用于多肽疫苗设计

Genetic analysis, techniques and applications Pub Date : 1996-09-01 DOI: 10.1016/1050-3862(95)00156-5

Kamalakar Gulukota, Charles DeLisi

{"title":"HLA allele selection for designing peptide vaccines","authors":"Kamalakar Gulukota, Charles DeLisi","doi":"10.1016/1050-3862(95)00156-5","DOIUrl":"10.1016/1050-3862(95)00156-5","url":null,"abstract":"<div><p>A central problem in developing vaccines against rapidly evolving viruses such as HIV and Influenza is the mutability of their antigens. In principle, the problem can be mitigated by using peptides from conserved portions of viral proteins. However, because cytotoxic T lymphocytes (CTLs), which such vaccines would stimulate, recognize pathogenic peptides only in association with class I products of the Major Histocompatibility Complex (MHC), and because human leukocyte antigen genes (HLA; the human MHC) are highly polymorphic, a peptide vaccine would have to bind a number of different HLA products. A natural question then, which is pertinent to the safety of the vaccine is, which HLA molecules should be targeted to achieve a prespecified coverage (say 90%) of a population. Taking account of disequilibrium between linked HLA loci, we identify 3–6 class I HLA alleles, depending on ethnic group, which cover about 90% of the population. While this leaves large numbers of individuals uncovered, a high level of herd immunity, and hence eradication of the virus, can be achieved through such a vaccine.</p></div>","PeriodicalId":77142,"journal":{"name":"Genetic analysis, techniques and applications","volume":"13 3","pages":"Pages 81-86"},"PeriodicalIF":0.0,"publicationDate":"1996-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/1050-3862(95)00156-5","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19896087","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 46

Allele-specific hybridization of lipoprotein lipase and factor-V Leiden missense mutations with direct label alkaline phosphatase-conjugated oligonucleotide probes 用直接标记碱性磷酸酶偶联寡核苷酸探针对脂蛋白脂肪酶和因子- v Leiden错义突变进行等位基因特异性杂交

Genetic analysis, techniques and applications Pub Date : 1996-09-01 DOI: 10.1016/1050-3862(95)00149-2

Calvin P.H. Vary, Marybeth Carmody, Renee LeBlanc, Tim Hayes, Clark Rundell, Len Keilson

{"title":"Allele-specific hybridization of lipoprotein lipase and factor-V Leiden missense mutations with direct label alkaline phosphatase-conjugated oligonucleotide probes","authors":"Calvin P.H. Vary, Marybeth Carmody, Renee LeBlanc, Tim Hayes, Clark Rundell, Len Keilson","doi":"10.1016/1050-3862(95)00149-2","DOIUrl":"10.1016/1050-3862(95)00149-2","url":null,"abstract":"<div><p>Direct label alkaline phosphatase (AP) conjugated oligonucleotide probes (AP-DNA) were prepared to assess their utility for allele-specific detection of single base substitutions. Oligonucleotide conjugates were designed to detect point mutations in the genes for lipoprotein lipase (LPL) and coagulation factor-V (FV). Genomic DNA samples, including ones known to harbor point mutations in the genes for LPL and FV, were prepared from whole blood and subjected to polymerase chain reaction (PCR). PCR products were analyzed by Southern hybridization with the allele-specific AP-DNA probes and restriction endonuclease analysis. Thermal profiles for hybridization indicate optimal allele-specific selectivity was achieved with temperatures ranging from 45°C to 55°C at a total Na<sup>+</sup> concentration of 150 mM. Under these conditions the base changes studied were easily discriminated with allele specific hybridization signals in excess of 200:1 as estimated by scanning densitometry. Complete concordance was observed between hybridization and restriction analyses for 175 LPL and 201 FV clinical and reference samples. The total time for analysis of the PCR products was less than 2 h with a dot blot hybridization protocol.</p></div>","PeriodicalId":77142,"journal":{"name":"Genetic analysis, techniques and applications","volume":"13 3","pages":"Pages 59-65"},"PeriodicalIF":0.0,"publicationDate":"1996-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/1050-3862(95)00149-2","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"19896790","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 7