In summary , we applied rd - pcr technique to the isolation of cdna fragments , which , compared with the conventional cloning procedure , is more speedy and higher throughput . the rd - pcr cdna fragments were constructed into a s . cerevisiae est ( expressed sequence tags ) library , which lay the foundation of cdna probe preparation and preparation of cdna microarray 综上所述，本研究采用rd ? pcr技术，探索了利用该技术快速分离大量cdna片段的具体过程，并建立了酵母cdna文库及酵母cdna探针文库，为dna芯片的制备打好基础。
Constructing cdna expressing library of erythrocytic plasmodium falciparum from hainan : the total rna was obtained by using . triplix kit . a modified oligo ( dt ) primer ( cds ffi pcr primer ) was used in the single - stranded ( ss ) dna synthesis reaction . the ss - dna was reversely transcripted from total rna . double - stranded ( ds ) cdna was amplified by long - distance ( ld ) pcrafter the digestion with proteinase k and sfi i , the cdna with no less than 200bp was collected and purified by glass - milk kitthe library was constructed after the ligation of cdna to tiplex2 phage particle packaged with the packaging extract system in vitro . a high titer and high recombinant ratio of cdna library was constructed 构建恶性疟原虫海南株红内期cdna表达文库提取红内期恶性疟原虫海南株总rna ，直接以总rna为模板使用cdna文库构建试剂盒，首先反转录合成ss一dna ，再扩增合成ds一dna ( cdna ) ，对扩增产物用蛋白酶k消化及左z丁i酶切，抽提蛋白、去除rna后，用玻璃奶试剂盒纯化、回收20obp以上的片断，经与载体连接再用蛋白包装物包装后形成未扩增文库，最后扩增完成恶性疟原虫海南株红内期cdna表达文库的构建。
Part 1 screening of genes related to the ovary development of the mitten crab ( eriocheir japonica sinensis ) chapter 1 construction of subtractive cdna library of mitten crab ( eriocheir japonica sinensis ) ovary two subtracted cdna libraries of mitten crab ( eriocheir japonica sinensis ) ovaries at two successive developmental stages were constructed by suppression subtractive hybridization ( ssh ) 第一部分中华绒螯蟹卵巢发育相关基因的筛选1中华绒螯蟹卵巢差减cdna文库的构建应用抑制性差减杂交技术，构建了中华绒螯蟹卵巢两个发育时期的差减cdna文库。
A cdna subtractive library with high subtractive efficiency of repeated + gz exposures in rat brain was constructed with suppression subtractive hybridization ( ssh ) . the cdna subtractive library after amplification included 100 blue clones and 400 white clones , 75 ones of which were selected to prepare for plasmid . identification of the clones with restriction endonuclease cleavage showed most of them had been cloned to the vector 构建高消减效率的+ gz重复暴露大鼠脑cdna消减文库，扩增后cdna文库包含约400个白色克隆和100个蓝色克隆，克隆饱满清晰，随机挑取75个白色克隆，制备质粒后，以ecor酶切分析，表明大部分克隆入质粒载体。
After careful studying their relative importance to immune response and the possibility of the match , seventeen sequences of interest were selected for further experiment , including estss analogous to 11 . 5kd antibacterial peptide , lysozyme , serine protease and its inhibitor , lectin , antifreeze protein , et al . primers designed according to the sequences were used to amplify the corresponding estss from both blood and cephalothorax cdna library 在仔细分析了它们在免疫系统中的重要性和在对虾中出现的可能性之后，从中选出了17条可能编码抗菌肽，溶菌酶，凝集素、丝氨酸蛋白酶及其抑制剂，抗冻蛋白等蛋白质的序列，以此为依据设计引物，在中国对虾的血液和头胸部cdna文库中扩增相应的序列。
These results are very important for us to further understand the genetic background , biological characteristics , evolutionary rule and the anti - schistosomajaponicum mechanism of microtus fortis at the molecular levels . the specific base changes of the dna fragme nt between the two subspecies are probably correlative with the animal immigration , survival conditions , and species evolution . the cdna library of microtus fortis bone marrow cells was transferred in situ to nylon membrane , which was divided into eight equals ( ga ~ - gh ) 利用已经建立的东方田鼠骨髓细胞质粒cdna文库，将cdna文库转化菌落印迹至尼龙膜，将膜均分成8份( ga gh ) ，制备基因池，分别培养、提取基因池质粒dna ，通过lipofect - 2000脂质体转染技术，将基因池质粒dna导入hek293细胞， 48h后收集转染细胞上清液，即条件培养基。
It has made the strong basis for further studying mechanism of replication , virulence and determinant , attenuation , pathogenesis , functions of genetic products , specific diagnosis , cell and host tropism , development of dna vaccine and marker vaccine of csfv , and provided an excellent tool for molecular virology . main research contents include : based on published nucleotide sequences of csfv and by the help of computer analysis software , high conservative regions and single restriction enzyme sites of genome were selected . utilizing rt - pcr and nested - pcr techniques , 7 overlapping cdna fragments covering the full genome of csfv c - strain were successfully amplified 中国猪瘟兔化毒(脾淋毒)基因组cdna文库的构建、序列分析：根据已发表的猪瘟病毒( csfv )核苷酸序列，借助计算机软件分析，选择高保守区段和基因组中的单一限制性酶切位点，利用rt - pcr及nested - pcr和helf - nestedpcr技术，成功地扩增出了覆盖c -株全基因组的7个cdna重叠片段f1 f7 ，分别克隆到pmd - 18t或pgem - teasy载体进行测序后，拼接出了其核苷酸序列。
Positive probe was made from the floral meristem materials cultured for 5 days and 10 days with hormones by rna reverse transcription and labelling with radioactive dctp ; two negative probes from uncultured explants and it cultured for 5 days and 10 days . then , we did calculating signal arrays , sequencing , sequence analysis and alignments on genebank etc . finally 229 different ests were got from the cdna library 用在含有外源激素的培养基上培养5天(花分生组织开始形成)和10天(花分生组织已基本形成)的风信子外植体为材料，构建起cdna文库。利用cdnamicroarrray技术，经过杂交筛选和est分析序列分析，最终从cdna文库中获得了229个不同的外源激素诱导响应的基因序列。
( 3 ) 490bp cdna fragment of fe ( ii ) - transporter gene mxlrtl was cloned by using common pcr method from iron - stressed root cdna library of malus xiaojinensis cheng et jiang with primers designed according to the conserved domain sequences of plant irt gene families . then we cloned the full cdna of mxlrtl gene by race method from this library with primers designed according to the sequence of 490bp cdna fragment ( 3 )根据植物irt基因家族的功能保守区序列设计引物，首先通过常规pcr法从缺铁胁迫处理的小金海棠根系cdna文库中克隆了fe ( ) -转运蛋白基因mxirt1的490bp的片段，然后根据测序结果设计引物，通过race法从该cdna文库中克隆了mxirt1基因的cdna全长。
Furthermore , suppression subtractive hybridization ( ssh ) was employed for the isolation of cdna fragments for euonymus japonicus " zhuzi " differentially expressed genes , and forward suppression subtractive cdna library of cold - regulated genes was constructed . the seedlings of euonymus japonicus " zhuzi " treated with low temperature were as tester and untreated seedlings as driver . subtractive cdna library was differentially screened through cdna macroarray , six hundreds and four cdna clones were identified as cold specifically induced or highly expressed ( 5 )应用抑制差减杂交( suppressionsubtractivehybridization , ssh )方法，构建冷诱导表达的正向抑制差减cdna文库，低温处理的幼苗为tester ，常温处理为driver ，通过cdna微阵列差异筛选cdna文库，得到604个低温诱导或表达增强的候选克隆，对其中的84克隆进行dna测序，去除冗余的cdna ，在genbank中进行核酸和蛋白质同源性的比较和功能分析，共有36个单一序列，其中12个cdna在genbank数据库没有同源的序列。
