共查询到20条相似文献,搜索用时 140 毫秒
1.
2.
3.
利用基因芯片技术筛选HIV-1F亚型基因限制性显示探针 总被引:2,自引:0,他引:2
为筛选限制性显示技术制备的HIV 1F亚型基因探针 ,应用基因芯片打印仪将其有序地打印在玻片上制备基因芯片 .在随机引物延伸的过程中进行HIV样品的荧光标记 ,然后与芯片进行杂交 .杂交后清洗玻片并干燥 ,对芯片进行扫描 ,分析各探针的杂交信号 .从中筛选了 14个基因片段作为芯片下一步研究的探针 .实验证明 ,限制性显示技术是一种制备基因芯片探针的实用方法 相似文献
4.
5.
6.
建立制备炭疽芽胞杆菌检测基因芯片的技术,并探讨研制检测炭疽芽胞杆菌基因芯片的方法。酶切炭疽芽胞杆菌的毒素质粒和荚膜质粒,通过建立质粒DNA文库的方法获取探针,并打印在经过氨基化修饰的玻片上,制成用于炭疽芽胞杆菌检测的基因芯片。收集了290个阳性克隆探针,制备了检测炭疽芽胞杆菌的基因芯片。提取炭疽芽胞杆菌质粒DNA与基因芯片杂交,经ScanArray Lite芯片阅读仪扫描得到初步的杂交荧光图像。通过分析探针的杂交信号初步筛选出273个基因片段作为芯片下一步研究的探针。 相似文献
7.
探讨利用eDNA文库法制备HCV-1诊断基因芯片探针的可行性.用限制性内切酶Sau3AI消化HCVla及lb全长eDNA,所得的酶切片段72℃补平加A,AT克隆,PCR初步鉴定,并测序.结果显示:HCV两个亚型1a、1b的全长eDNA得到57个大小相对一致(200-1000bp)的片段,平均每个亚型约28个,PCR及序列分析表明,所扩增的片段均属于HCV-1的特异基因,可做为HCV-1诊断基因芯片探针.利用eDNA文库法收集片段是一种快速、简便制备芯片探针的实用方法. 相似文献
8.
基因芯片研究中,基于不同的样品来源,基因含量,检测方法和分析目的,采用的核酸分离。扩增和标记方法各异。本综述了对核酸样品制备条件的优化,主要有核酸的单链化处理,片段化和标记方法,根据具体情况选用合适的处理方法,可显提高基因芯片检测的特异性和重现性。 相似文献
9.
《生物技术通讯》2020,(2)
分子诊断技术主要包括核酸分子杂交技术、核酸扩增技术、基因芯片技术、基因测序技术,已广泛应用于临床各科。比如在肿瘤疾病中应用荧光原位杂交技术(FISH)检测乳腺癌中HER-2基因扩增,应用基因芯片技术检测胃癌进程中基因拷贝数变化,应用高通量测序技术(HTS)检测肺癌中的EGFR基因突变;在遗传病中应用FISH技术针对唐氏综合征进行产前诊断,应用数字PCR技术进行无创产前筛查,应用HTS技术进行Hermansky-Pudlak综合征的诊断;在感染性疾病中应用RT-PCR试剂盒进行新冠肺炎的检测,应用基因芯片技术筛选抗生素抗性基因,应用HTS技术进行耐药基因的筛选等。本文主要介绍分子诊断技术在这三大类疾病领域的临床应用最新进展。 相似文献
10.
11.
DNA芯片技术研究进展 总被引:66,自引:5,他引:61
DNA芯片技术是近年来发展迅速的生物高技术 .其基本过程是采用寡核苷酸原位合成或显微打印手段 ,将大量探针片段有序地固化于支持物如硅芯片的表面 ,然后与扩增、标记的生物样品杂交 ,通过对杂交信号的检测分析 ,即可得出样品的遗传信息 .该技术不仅可以对遗传信息进行定性、定量分析 ,而且扩展到基因组研究和基因诊断等方面的应用 .尽管目前在硬件和软件上还面临一些困难 ,但其发展和应用的前景广阔 . 相似文献
12.
对流感病毒14个血凝素亚型的基因芯片检测技术进行了初步研究。通过RT-PCR克隆禽流感病毒血凝素基因片段,获得重组质粒。从重组质粒扩增大约500bp的DNA片段,浓缩后点到氨基化玻璃载体上,制成芯片。待检病毒样品用TRIzolLS提取RNA,反转录过程中用Cy5标记样品cDNAs。将标记样品与芯片杂交,扫描芯片上待检样品与芯片上捕捉探针的结合位点,杂交信号与预期设想一致。结果显示,DNA芯片技术可以提供一种有效的AIV血凝素亚型鉴别诊断方法。 相似文献
13.
Pelizzari CA Khodarev NN Gupta N Calvin DP Weichselbaum RR 《Nucleic acids research》2000,28(22):4577-4581
DNA arrays and chips are powerful new tools for gene expression profiling. Current arrays contain hundreds or thousands of probes and large scale sequencing and screening projects will likely lead to the creation of global genomic arrays. DNA arrays and chips will be key in understanding how genes respond to specific changes of environment and will also greatly assist in drug discovery and molecular diagnostics. To facilitate widespread realization of the quantitative potential of this approach, we have designed procedures and software which facilitate analysis of autoradiography films with accuracy comparable to phosphorimaging devices. Algorithms designed for analysis of DNA array autoradiographs incorporate 3-D peak fitting of features on films and estimation of local backgrounds. This software has a flexible grid geometry and can be applied to different types of DNA arrays, including custom arrays. 相似文献
14.
D Guiliano M Ganatra J Ware J Parrot J Daub L Moran H Brennecke J M Foster T Supali M Blaxter A L Scott S A Williams B E Slatko 《BioTechniques》1999,27(1):146-152
A chemiluminescent approach for sequential DNA hybridizations to high-density filter arrays of cDNAs, using a biotin-based random priming method followed by a streptavidin/alkaline phosphatase/CDP-Star detection protocol, is presented. The method has been applied to the Brugia malayi genome project, wherein cDNA libraries, cosmid and bacterial artificial chromosome (BAC) libraries have been gridded at high density onto nylon filters for subsequent analysis by hybridization. Individual probes and pools of rRNA probes, ribosomal protein probes and expressed sequence tag probes show correct specificity and high signal-to-noise ratios even after ten rounds of hybridization, detection, stripping of the probes from the membranes and rehybridization with additional probe sets. This approach provides a subtraction method that leads to a reduction in redundant DNA sequencing, thus increasing the rate of novel gene discovery. The method is also applicable for detecting target sequences, which are present in one or only a few copies per cell; it has proven useful for physical mapping of BAC and cosmid high-density filter arrays, wherein multiple probes have been hybridized at one time (multiplexed) and subsequently "deplexed" into individual components for specific probe localizations. 相似文献
15.
Lubing Zhou Yuan-Di Halvorsen Ellen V. Cryan Patricia D. Pelton Thomas P. Burris Keith T. Demarest 《Biotechnology Techniques》1999,13(8):513-517
High density DNA microarrays containing over 5000 cDNA clones were used to carry out a comprehensive investigation of gene expression during adipogenesis. Complex probes synthesized from total RNA were hybridized to the arrays to determine the level of mRNA expression of each arrayed gene. Thirty three genes (29 known and 4 ESTs with no identified homologies) have been found to alter their level of expression more than 2.5-fold after differentiation. The quantitative measurement by DNA array was in good agreement with conventional Northern blot analysis of selected genes. Our results demonstrate that utilization of a DNA array is a speedy, efficient and quantitative approach to profile the expression of a large number of genes. 相似文献
16.
17.
用基因表达谱芯片研究人正常肝和肝细胞癌中差异表达的基因 总被引:35,自引:0,他引:35
以基因表达谱芯片对人正常肝及肝癌组织基因表达的差异性进行了研究比较。奖4096条人cDNA用点样仪点在特制玻片上制备成表达谱芯片;利用肝和肝癌组织的mRNA通过逆转录方法,将Cy3和Cy52种荧光分别标记到两种组织的cDNA上,制备成cDNA探针,并与表达谱芯片进行杂交及扫描,重复4次实验,通过计算机数据处理判定基因是否在上述2种组织中有表达差异,筛选出差异表达的基因共903条。基因芯片技术可同时 相似文献
18.
Zhuo ZHOU Zhi-xun DOU Chen ZHANG Hou-qing YU Yi-jie LIU Cui-zhu ZHANG You-jia CAO 《Virologica Sinica》2007,22(4):326-335
DNA microarrays have been acknowledged to represent a promising approach for the detection of viral pathogens. However, the probes designed for current arrays could cover only part of the given viral variants, that could result in false-negative or ambiguous data. If all the variants are to be covered, the requirement for more probes would render much higher spot density and thus higher cost of the arrays. Here we have developed a new strategy for oligonucleotide probe design. Using type I human immunodeficiency virus (HIV-1) tat gene as an example, we designed the array probes and validated the optimized parameters in silico. Results show that the oligo number is significantly reduced comparing with the existing methods, while specificity and hybridization efficiency remain intact. The adoption of this method in reducing the oligo numbers could increase the detection capacity for DNA microarrays, and would significantly lower the manufacturing cost for making array chips. 相似文献
19.