1.Methods:Through looking up bioinformatics,we chose ATPIF1 as candidate gene and uesd primer3 to design its primer. Mutation analysis was carried out by polymerse chain reaction(PCR) and DAN direct secquencing.
方法:引物设计应用在线引物设计软件—Primer 3,采取PCR扩增,直接测序的方法进行候选基因ATPIF1的突变分析。
2.Diagnosis of CPV infection should be based on the study of P32 protein. Pairs of primers were designed with Primer 5.0 according to Goat pox virus (GTPV) P32 sequence published in Genbank.
本实验根据GenBank中发表的山羊痘病毒(Goat pox virus,GTPV)P32基因序列,利用软件Primer 5.0分别设计了P32鉴定和表达用引物。
3.First , the Oligo dT-3sites Adaptor primer was used to conduct reverse transcription.
首先，以试剂盒提供的Oligo dT-3sites Adaptor primer为引物进行逆转录；
4.RAPD reaction system of peach was Optimized:Total volume was 25 μl,including of 40ng template DNA, 2.0mM MgCl_2, 150μM dNTP, 2.5μl 10× buffer, 1U Taq DNA polymerase, 20pmol Primer. the rest was supplied with ddH_2O.
优化了桃适宜的RAPD反应体系：总反应体积为25μl，其中模板DNA40ng，MgCl_22.0mM，dNTP 150μM，10×buffer 2.5μl，Taq DNA聚合酶lU，Primer20pmol，余下的用ddH_2O补充。
5.According to the 16S rRNA gene sequence published in GenBank, which include Mycoplasmas gallisepticum of chicken and M.hyopneumoniae,M. hyosynoviae and M.flocculare of swine(submitted No.:Mg AY744942, Mhp AE017244, Mhs AY973563 and Mf X63377), and the PCR primers labled by Dig and the probe labled by biotin were designed by the software DNAstar and Primer 5.0. The system and condition of PCR-ELISA were optimized.
实验依据GenBank中登录的鸡源支原体和猪源支原体16SrRNA基因序列(登录号:Mg,AY744942;Mhp,AE017244;Mhs,AY973563;Mf,X63377),运用DNAStar软件和Primer5.0软件设计PCR特异性引物和探针,其中引物用地高辛标记,探针用生物素标记。
6.METHODS: The genotypes of 90 Chinese Han in Guangdong and 104 Sherpas in Tibet were analyzed by sequence special primer polymerase chain reaction(SSP-PCR) sequencing the surfactant protein A gene.
方法:应用序列特异性引物-聚合酶链反应(polymerase chain reac-tion-sequence specific primer,SSP-PCR)方法,对90名广东汉族人和104例西藏夏尔巴人SP-A基因进行检测。
7.Apair of primers was designed to amplify VP2 gene by PCR according to the published sequence of CPV’s VP2 gene with primer5.0 software. The product of PCR named VP2 is approximate 1.8kb in length.
根据Genbank已发表的CPV-2 VP2 基因的序列,利用primer 5.0 软件设计并合成一对引物,通过PCR 扩增出长约1.8kb 的基因片段,命名为VP2。
8.By analyzing the differential gene fragments in the chrysanthemum and in the Arabidopsis thaliana, it is found that SA5-2 in the chrysanthemum and SA3 in the Arabidopsis thaliana are very similar(Fig 5.13,5.14), which are amplified from primer combination with the same anchored primer: AAG CTT TTT TTT TTA and the similar random primer. The sequence of random primer is AAGCTTAGTAGGC and AAGCTTTGGTCGA respectively.
分析菊花和拟南芥经声波处理后产生的差异条带。 经比较发现，菊花中的特异片段SA5-2和拟南芥中的特异片段SA3(见图5．13，5．14)有相似之处：扩增这两条条带的引物组合相近，锚定引物均为AAG CTT TTT TTT TTA，随机引物分别为AAGCTTAGTAGGC和AAGCTTTGGTCGA；
9.Conclusions The universal primer(CHS1 1S,CHS1 1R)is regarded as a specificity primer. The result showed that the sensitivity of universal primer PCR is 100 fg.
结论皮肤病原真菌通用引物(CHS1 1S,CHS1 1R)具有较强的特异性,敏感度为100 fg;
10.Each primer amplified three to sixteen bands. The number of bands amplified by the same primer (except for OPM 19 primer) is different in the two species, and their type of bands is clearly different.
每一引物的扩增带数为 3～ 16条 ,同一引物对两种贝类的扩增带数不同 (OPM19除外 ) ,对两贝类的扩增带型也有较大差异。
