Northern杂交

摘要： 为了给短小芽孢杆菌的改造提供新思路,提高碱性蛋白酶产量,实验室前期对短小芽孢杆菌进行了转录组测序,预测了短小芽孢杆菌的sRNA.本研究通过生物信息学方法和Northern杂交鉴定了一个新的sRNA Bpsr112.然后构建了Bpsr112的敲除型菌株和过表达菌株,利用相关菌株进行了生长曲线、盐胁迫和蛋白酶活等实验,再利用SDS-PAGE检测胞外蛋白酶AprE的表达水平.结果 表明,在发酵至60 h和72 h时,与对照菌株相比,敲除型菌株酶活显著降低(P<0.01),过表达菌株酶活显著提高(P<0.01);同时SDS-PAGE结果表明,AprE蛋白含量在敲除菌株中降低,在过表达菌株中提高,说明sRNA Bpsr112对短小芽孢杆菌蛋白酶活具有正调控.本研究鉴定了短小芽孢杆菌中的sRNA,并发现Bpsr112对蛋白酶活具有促进作用.

摘要： In order to establish a more sensitive and specific molecular hybridization detection system of Tobacco mosaic virus Liaoning isolate(TMV-LN),total viral RNA was extracted from purified TMV-LN viral particles and amplified by reverse-transcript PCR. The PCR products were ligated into pUC119 vector to construct pUCTMV-PP that express RNA detection probe using Digoxigenin(DIG)Northern Starter Kit. DNA probe was also constructed using DIG DNA hybridization kit. Dot-blot hybridization and Northern blot hybridization analysis were performed to study the specificity and sensitivity of the RNA probe and DNA probe constructed above. Both of the probes showed high specificity and sensitivity in TMV-LN detection through Dot-blot hybridization and Northern blot hybridization detection system. Comparison of two hybridization systems,Dot-blot hybridization system is suitable for virus qualitative detection,whereas,Northern blot hybridization has its advantage in relative quantification of viral genome RNA.%为了建立烟草花叶病毒辽宁分离物(TMV-LN)的高特异性、高灵敏度的分子杂交检测体系,从粗提纯的TMV-LN粒子中提取RNA,设计特异性引物通过RT-PCR扩增TMV-LN的CP和3'端非翻译区域,将片段连至pUC119载体获得重组质粒pUCTMV-PP,体外转录获得地高辛(DIG)标记的TMV-LN正义链RNA杂交检测探针,同时构建DNA检测探针作为对照.采用点印迹(Dot-blot)杂交和Northern杂交对比RNA探针和DNA探针对TMV-LN的检测特异性和灵敏性.检测结果表明,RNA探针和DNA探针在点印记杂交和Northern杂交中均表现出良好的检测特异性,RNA探针在检测灵敏度方面要略好于DNA探针,且点印迹杂交体系在病毒定性方面较为快捷,Northern杂交体系在病毒基因组RNA的定量方面具有明显优势.

摘要： 为了探讨蛹虫草类枯草杆菌蛋白酶（Subtilisin-like protease）的表达特性，以真菌蛹虫草菌丝体为研究对象，通过 RT-PCR 获得蛹虫草 CmKexin 基因的 ORF 序列，并进行序列分析。应用 Northern 杂交和 Real-time PCR 方法，检测蓝光照射后蛹虫草菌丝体中 CmKexin 基因的转录情况。结果获得蛹虫草 CmKexin 基因的 ORF 序列。对翻译蛋白质产物 CmKexin 进行生物信息学分析，表明该蛋白质含1个属蛋白转化酶的肽酶 S8家族功能域（143～429）和1个前体蛋白转化酶 P 功能域（514～600），符合真菌类枯草杆菌蛋白酶的特征。蛹虫草菌丝体在黑暗预培养4 d 后再蓝光照射，Northern Blotting 和 Real-time PCR 检测均可得到相同的结果，即在持续的蓝光照射48～50 h 时，出现 CmKexin 基因的瞬时大量转录。而其他时间内均未检测到该基因的大量转录。试验结果将为蛹虫草类枯草杆菌蛋白酶的利用提供理论依据。%In order to study the characteristics of subtilisin-like protease expression in the mycelia of fungus Cordyceps militaris,CmKexin gene ORF sequences of C.militaris was got by RT-PCR,then CmKexin sequence was analysed.The Real-time PCR and Northern Blotting were used to the CmKexin gene expression analysis in the C.militaris mycelia in blue light irradiation of different time.Results showed the CmKexin gene ORF sequence was obtained from C.militaris mycelia.The bioinformatics analysis for the translation products of CmKexin gene showed that there were 1 peptidase S8 family domain of protein convertases (143 -429)and 1 proprotein convertase P-do-main(5 14 -600)in the protein.The two domains existed in CmKexin demonstrate the characteristics of subtilisin-like protease.Under the dark pre-culture in 96 h,a large amounts of transcript of CmKexin gene in the mycelia could be detected by Northern Blotting and Real-time PCR in the continuous blue light irradiation to 48 -50 hours. But the transcription of CmKexin gene wasn′t detected in the other time of C.militaris mycelia culture.These results would provide the basis for using the subtilisin-like protease in Cordyceps militaris.

摘要： In order to study the molecular mechanism of the " albino-after green" in wheat ( Triticum aestivum L. ) albinism line, we have designed the PCR primer with the LSC of Chloroplasts genome sequence in Chinese spring wheat. We have amplified the related gene segment in chinese spring wheat, albinism line and Aibian 1 and get the specific gene fragment WFC01 in albinism line. The total RNA has been extracted from leaves in Chinese spring wheat, Aibian 1 and albinism line. WFC01 was the probe for northern blot. The results show that the expression of gene WFC01 has been inhibition along with the leaves albinism in albinism line compared with albinism line and Chinese spring wheat. The gene WFC01 expression was back to normal with the leaves get green.%为研究小麦返白系阶段性“白化-复绿”的分子机制,以中国春小麦叶绿体基因组序列的LSC区为参考序列,分段设计引物,PCR扩增中国春、矮变1号、返白系的相应基因片段,在返白系中获得特异基因片段WFC01.提取中国春、矮变1号幼嫩叶片和返白系白化及复绿叶片总RNA,毛细管法转膜,WFC01为探针进行Northern杂交.结果表明,与对照矮变1号和中国春相比,返白系随着叶片的白化,WFC01基因片段的表达受到明显抑制,几乎无表达,随着叶片的复绿,WFC01基因片段的表达恢复正常,与对照表达量一致,表明该基因的表达与返白系的阶段性白化、复绿有关.

摘要： [Objective] The aim of this study is to clone SCARECROW-LIKE 7 (SCL7) gene,to analyze its molecular mechanisms and promote their applications in breeding.[Method] The total RNA from the leaves of tobacco was used as the template to design the degenerate primers based on homology cloning strategy,and then the full-length cDNA sequence ofzmSCL7 was obtained through a combined reverse transcription-PCR (RT-PCR).Bioinformatics method was used to analyze the sequence characteristics of this gene.Northem blot was used to investigate the expression pattern.Western blot was used to investigate the transgenic analysis.Chlorophyll content,Fv/Fm,MDA and proline content were investigated for functional verification of salt resistance.[Result] The results indicated that cDNA of ZmSCL7 was 1 653 bp and contained a single open reading frame of 550 amino acid residues.Northern blot indicated that the mRNA accumulation of ZmSCL7 was induced by low temperatures,salt stress,abscisic acid (ABA) and H2O2.Additionally,compared with the control,transgenic ZmSCL7 tobacco,germination was inhibited less.Chlorophyll content,Fv/Fm,proline content increased and MDA content decreased.[Conclusion] ZmSCL 7,a transcription factor,was induced by a variety of stress and could increase salt tolerance by overexpression in tobacco.%[目的]对玉米SCARECROW-LIKE7(SCL7)进行克隆与表达研究,了解该基因表达的分子机制及其应用.[方法]以玉米叶片总RNA为模板,根据同源克隆策略设计简并引物,利用RT-PCR结合RACE技术,获得ZmSCL7的全长cDNA序列.利用同源性比对进行序列分析,通过Northern杂交分析ZmSCL7在不同逆境胁迫下的表达特征,对转基因烟草进行Western blot分析,并测定最大光化学效率、叶绿素、丙二醛和脯氨酸含量验证该基因的抗盐功能.[结果]获得ZmSCL7全长cDNA序列1653 bp,编码550个氨基酸.Northern杂交表明该基因在NaC1、H2O2和低温处理条件下上调表达,在ABA处理条件下下调表达.与对照相比,转ZmSCL7烟草在200mmol·L-1NaCl时萌发受到较小抑制,且最大光化学效率、叶绿素和脯氨酸含量上升,丙二醛含量下降.[结论]ZmSCL7作为一个转录因子受多种逆境胁迫诱导,该基因的过量表达提高了烟草的抗盐性.

摘要： [目的]确定真菌Mucor amphibiorum RCS1中一个类S-腺苷-L-高半胱氨酸水解酶基因(S-adenosyl-L-homocysteine hydrolase-like,sahhl)受蓝光诱导表达.[方法]以真菌M.amphibiorum RCS1为研究对象,在随机PCR过程中从中获得了一段555 bp长度的DNA序列.以地高辛对这段已知序列进行标记制备探针,通过Northern杂交检测M.amphibiorum RCS1菌丝体培养过程中,由黑暗到蓝光再到黑暗这一光照条件改变的情况下,sahhl基因的转录情况.同时结合应用real-time PCR方法进行分析检测.[结果]经过比对确定这段555bp序列与已经发表的人(Homo sapiens)、家鼠(Mus musculus)和部分真菌的,S-腺苷-L-高半胱氨酸水解酶基因sahh有较高的同源性.因此,初步确认这段mRNA序列来自M.amphibiorum RCS1的一个类S-腺苷-L-高半胱氨酸水解酶基因.sahhl基因在黑暗预培养24 h的情况下,蓝光诱导24 h时通过Northern杂交和real-time PCR均可从菌丝体中检测到sahhl基因的大量转录.但sahhl基因在黑暗预培养48 h的情况下,通过real-time PCR没有检测到sahhl基因的大量表达.[结论]上述结果说明,蓝光可以诱导M.amphibiorum RCS1中生长旺盛的菌丝体中sahhl基因的表达.

摘要： The sugar beet buds treated with 4 °C were tested and the sugar beet buds treated with 25 °C were used as the control. Ty7Br600 gene was randomly labeled with a-32P-ATP, and Northern blotting was conducted. Northern blotting results showed that Ty7Br600 gene expressed in sugar beet buds treated with 4 °C for 72 d, but it's expression could not be detected in the control treated with 25°C. Ty7Br600 could be a new gene related with biennial sugar beet boltting, which was induced by low temperature. Southern blotting results showed that there were 2-3 copy numbers in the genomic DNA, which suggested that Ty7Br600 should be a gene fragment of sugar beet genome, and have 2 or low copies in sugar beet genome.%以低温诱导甜菜幼苗为试验材料,以来进行低温诱导的甜菜幼苗为对照,采用α-32P-ATP放射标记按随机引物标记法标记本实验室克隆的TyBr600基因,进行Northern杂交.Northern杂交试验结果显示,在低温诱导培养的甜菜幼苗的RNA群体中,出现较强的杂交条带,而在未诱导培养的甜菜幼苗的RNA群体中,则几乎没有阳性杂交条带出现.因此,TyBr600这一序列有可能是二年生甜菜幼苗经低温诱导处理之后被诱导表达的与抽薹相关的新基因序列.Southern杂交结果发现,在每一组酶切中至少有2～3条条带,表明Ty7Br600确为甜菜基因组片段,也同时表明该基因在甜菜基因组中以2个拷贝或低拷贝形式存在.

摘要： 温度逆境胁迫可以诱导热激蛋白的表达,而热激处理能够减轻果实冷害。为了探讨温度逆境对香蕉热激蛋白基因表达的调控、辨析香蕉热激蛋白基因的表达与果实冷害的关系,采用同源序列法分离了香蕉果皮HSP70基因序列-根据已经报道的HSP70基因的氨基酸保守序列设计引物、以香蕉果皮总RNA为模板,用RT-PCR方法克隆香蕉的HSP70 cDNA,Northern杂交分析该基因在不同贮藏温度下的表达特征。结果得到2个序列不同的HSP70基因片段,分别命名为Ma-HSP70-1和Ma-HSP70-2,Ma-HSP70-1和Ma-HSP70-2之间的碱基同源性为86.9%,氨基酸同源性为97.1%。Northern杂交的结果表明,38°C短期热激处理诱导了Ma-HSP70-2基因的表达,低温冷藏能使2个基因的表达增强。并初步认为Ma-HSP70可能与香蕉果实耐冷性有关。

摘要： AIM: To investigate whether DNA-dependent activator of interferon-regulatory factors (DAI) inhibits hepatitis B virus (HBV) replication and what the mechanism is. METHODS: After the human hepatoma cell line Huh7 was cotransfected with DAI and HBV expressing plasmid, viral protein (HBV surface antigen and HBV e antigen) secretion was detected by enzyme-linked immunosorbent assay, and HBV RNA was analyzed by real-time polymerase chain reaction and Northern blotting, and viral DNA replicative intermediates were examined by Southern blotting. Interferon regulatory factor 3 (IRF3) phosphorylation and nuclear translocation were analyzed via Western blotting and immunofluorescence staining respectively. Nuclear factor-B (NF- B) activity induced by DAI was detected by immunofluorescence staining of P65 and dual luciferase reporter assay. Transwell co-culture experiment was performed in order to investigate whether the antiviral effects of DAI were dependent on the secreted cytokines. RESULTS: Viral protein secretion was significantly reduced by 57% (P < 0.05), and the level of total HBV RNA was reduced by 67% (P < 0.05). The viral core particle-associated DNA was also dramatically down-regulated in DAI-expressing Huh7 cells. Analysis of involved signaling pathways revealed that activation of NF-B signaling was essential for DAI to elicit antiviral response in Huh7 cells. When the NF-B signaling pathway was blocked by a NF-B signaling suppressor (I B -SR), the anti-HBV activity of DAI was remarkably abrogated. The inhibitory effect of DAI was independent of IRF3 signaling and secreted cytokines. CONCLUSION: This study demonstrates that DAI can inhibit HBV replication and the inhibitory effect is associated with activation of NF-B but independent of IRF3 and secreted cytokines.

摘要： Eight 35SP-rolB transgenic lines of T7 generation of upland cotton were applied to analyze the stabilities of transgenice inheritance and transcription. Total DNA and RNA were prepared from the leaf extracts of the rolB transgenic lines and the control, and the result was verified by PCR amplification and Northern hybridization. The results showed that the 35SP-rolB gene and the NPT Ⅱ selection marker in the genomes of transgenic lines, which were selected for seven successive generations based on the specific traits of strong rooting ability, decrepitude resistance and rapid recovery after seedling transplantation, behaved stable inheritance and transcription in all eight transgenic lines. The results also demonstrated that the orientated selection at toward target traits is an efficient way in keeping the genetic stability of the transgenic lines and the expressional stability of the 35SP-rolB transgene.%以8个T7代的棉花35SP-rolB转基因系为材料,通过提取转基因株系及对照植株叶片总DNA和总RNA,对各个转基因株系分别进行PCR检测和Northern杂交,研究转基因的遗传稳定性和转录稳定性.结果表明,35SP-rolB目的基因和NPT Ⅱ选择标记基因在经过对转基因系的连续多代定向选择后,在各个转基因系基因组中都能够稳定遗传和高效转录,说明针对强生根、早发、抗早衰等特殊性状进行定向选择是保持rolB转基因系遗传稳定性和转基因表达稳定性的有效途径.

摘要： 以8个T7代的35SP-rolB棉花转基因系为材料,对转基因的遗传稳定性和转录稳定性进行了分析.PCR检测和Northern杂交结果表明,35SP-rolB目的基因和NPTⅡ选择标记基因在经过对转基因系的连续多代定向选择后,在各个转基因系基因组中都能够稳定遗传和高效转录,说明针对强生根、早发、抗早衰等特殊性状进行定向选择是保持rolB转基因系遗传稳定性和转基因表达稳定性的有效途径.

摘要： 以8个T7代的35SP-rolB棉花转基因系为材料,对转基因的遗传稳定性和转录稳定性进行了分析.PCR检测和Northern杂交结果表明,35SP-rolB目的基因和NPTⅡ选择标记基因在经过对转基因系的连续多代定向选择后,在各个转基因系基因组中都能够稳定遗传和高效转录,说明针对强生根、早发、抗早衰等特殊性状进行定向选择是保持rolB转基因系遗传稳定性和转基因表达稳定性的有效途径.

摘要： 以8个T7代的35SP-rolB棉花转基因系为材料,对转基因的遗传稳定性和转录稳定性进行了分析.PCR检测和Northern杂交结果表明,35SP-rolB目的基因和NPTⅡ选择标记基因在经过对转基因系的连续多代定向选择后,在各个转基因系基因组中都能够稳定遗传和高效转录,说明针对强生根、早发、抗早衰等特殊性状进行定向选择是保持rolB转基因系遗传稳定性和转基因表达稳定性的有效途径.

摘要： 以8个T7代的35SP-rolB棉花转基因系为材料,对转基因的遗传稳定性和转录稳定性进行了分析.PCR检测和Northern杂交结果表明,35SP-rolB目的基因和NPTⅡ选择标记基因在经过对转基因系的连续多代定向选择后,在各个转基因系基因组中都能够稳定遗传和高效转录,说明针对强生根、早发、抗早衰等特殊性状进行定向选择是保持rolB转基因系遗传稳定性和转基因表达稳定性的有效途径.

摘要： 以8个T7代的35SP-rolB棉花转基因系为材料,对转基因的遗传稳定性和转录稳定性进行了分析.PCR检测和Northern杂交结果表明,35SP-rolB目的基因和NPTⅡ选择标记基因在经过对转基因系的连续多代定向选择后,在各个转基因系基因组中都能够稳定遗传和高效转录,说明针对强生根、早发、抗早衰等特殊性状进行定向选择是保持rolB转基因系遗传稳定性和转基因表达稳定性的有效途径.

摘要： 以8个T7代的35SP-rolB棉花转基因系为材料,对转基因的遗传稳定性和转录稳定性进行了分析.PCR检测和Northern杂交结果表明,35SP-rolB目的基因和NPTⅡ选择标记基因在经过对转基因系的连续多代定向选择后,在各个转基因系基因组中都能够稳定遗传和高效转录,说明针对强生根、早发、抗早衰等特殊性状进行定向选择是保持rolB转基因系遗传稳定性和转基因表达稳定性的有效途径.

摘要： 对经PK-15细胞培养的PCV2中国分离株优势基因型(CHN-2H)和非优势基因型(CHN-2A)进行了转录分析.结果可检测到四个RNA转录本,其中病毒复制酶蛋白基因RNA(Rep)、病毒核衣壳蛋白基因RNA(CR)和另外一个基因RNA(命名为AD)在接种细胞12h后用Northern杂交可检出;用RT-PCR技术可检测到全部转录本(包括另外一个新命名的Reps).同时结果揭示了在CHN-2H和CHN-2A之间转录本存在量和质的变化,表现为CHN-2A的Rep表达量大,CHN-2H的CR表达时间长和没有AD和Reps两个转录本.此研究为阐明PCV2流行的分子机制奠定了理论基础.

摘要： 对经PK-15细胞培养的PCV2中国分离株优势基因型(CHN-2H)和非优势基因型(CHN-2A)进行了转录分析.结果可检测到四个RNA转录本,其中病毒复制酶蛋白基因RNA(Rep)、病毒核衣壳蛋白基因RNA(CR)和另外一个基因RNA(命名为AD)在接种细胞12h后用Northern杂交可检出;用RT-PCR技术可检测到全部转录本(包括另外一个新命名的Reps).同时结果揭示了在CHN-2H和CHN-2A之间转录本存在量和质的变化,表现为CHN-2A的Rep表达量大,CHN-2H的CR表达时间长和没有AD和Reps两个转录本.此研究为阐明PCV2流行的分子机制奠定了理论基础.

摘要： 对经PK-15细胞培养的PCV2中国分离株优势基因型(CHN-2H)和非优势基因型(CHN-2A)进行了转录分析.结果可检测到四个RNA转录本,其中病毒复制酶蛋白基因RNA(Rep)、病毒核衣壳蛋白基因RNA(CR)和另外一个基因RNA(命名为AD)在接种细胞12h后用Northern杂交可检出;用RT-PCR技术可检测到全部转录本(包括另外一个新命名的Reps).同时结果揭示了在CHN-2H和CHN-2A之间转录本存在量和质的变化,表现为CHN-2A的Rep表达量大,CHN-2H的CR表达时间长和没有AD和Reps两个转录本.此研究为阐明PCV2流行的分子机制奠定了理论基础.

摘要： 对经PK-15细胞培养的PCV2中国分离株优势基因型(CHN-2H)和非优势基因型(CHN-2A)进行了转录分析.结果可检测到四个RNA转录本,其中病毒复制酶蛋白基因RNA(Rep)、病毒核衣壳蛋白基因RNA(CR)和另外一个基因RNA(命名为AD)在接种细胞12h后用Northern杂交可检出;用RT-PCR技术可检测到全部转录本(包括另外一个新命名的Reps).同时结果揭示了在CHN-2H和CHN-2A之间转录本存在量和质的变化,表现为CHN-2A的Rep表达量大,CHN-2H的CR表达时间长和没有AD和Reps两个转录本.此研究为阐明PCV2流行的分子机制奠定了理论基础.

摘要： 本发明公开了一种免预杂交的Northernblot杂交液及其应用，该免预杂交的Northernblot杂交液的pH值为7.4，包括如下终浓度的组分：5×SSC、20mMNa2HPO4、质量百分比为7％的SDS、质量百分比为0.02％的聚乙烯吡咯烷酮、质量百分比为0.02％的聚蔗糖400、质量百分比为0.02％的BSA、质量百分比为0.5％的吐温‑20；所述5×SSC含有0.75M的NaCl和0.075M的柠檬酸钠。采用本发明的Northernblot杂交液，无需进行杂交液与膜的预杂交步骤，即可直接将探针加入到杂交液中，使探针与尼龙膜上RNA进行杂交，减少实验步骤和时间。

摘要： 本发明公开了一种针对于不同GC％含量的基因芯片杂交组件。其包括至少两种探针和杂交溶液；其中，所述芯片上探针序列GC％含量范围在10％‑80％；所述杂交溶液包含样品片段、甜菜碱和Zn2+。利用本发明基因芯片杂交组件能够使不同GC％含量的DNA片段在同一个条件下进行杂交，并可消除非特异性杂交的信号。

摘要： 本发明提供了一种简单可靠的使样品生物聚合物与探针DNA相互杂交的方法，该方法不用盖玻片和密闭容器，而是在含有与含样品生物聚合物溶液蒸气压相同的溶液的密闭容器中只使含样品生物聚合物溶液与固定有探针生物聚合物的载玻片接触，从而使样品生物聚合物与探针生物聚合物相互杂交。还提供了用于本发明的杂交微阵列和试剂盒。

摘要： 本发明公开了一种免预杂交的Northernblot杂交液及其应用，该免预杂交的Northernblot杂交液的pH值为7.4，包括如下终浓度的组分：5×SSC、20mMNa2HPO4、质量百分比为7％的SDS、质量百分比为0.02％的聚乙烯吡咯烷酮、质量百分比为0.02％的聚蔗糖400、质量百分比为0.02％的BSA、质量百分比为0.5％的吐温‑20；所述5×SSC含有0.75M的NaCl和0.075M的柠檬酸钠。采用本发明的Northernblot杂交液，无需进行杂交液与膜的预杂交步骤，即可直接将探针加入到杂交液中，使探针与尼龙膜上RNA进行杂交，减少实验步骤和时间。

摘要： 本发明涉及植物病毒检测技术领域，尤其涉及一种基于Northern杂交检测番茄黄化曲叶病毒的方法。包括以下步骤：步骤1.根据番茄黄化曲叶病毒基因组序列设计引物C4‑F和C4‑R，PCR法制备带地高辛标记的番茄黄化曲叶病毒特异性DNA探针；步骤2.感染番茄黄化曲叶病毒3天后的烟草总RNA提取和总RNA在琼脂糖胶上的分离；步骤3.虹吸法转移步骤2中分离的总RNA至带正电的尼龙膜，转膜结束后在紫外交联仪下固定膜上的总RNA；步骤4.预杂交，并使用步骤1中制备的探针进行杂交；步骤5.洗涤杂交膜及杂交信号检测。该方法克服了现有检测技术的缺点，提供了一种灵敏、特异检测番茄黄化曲叶病毒的方法。

摘要： 本申请公开了一种基于Northern杂交检测家蚕microRNA的方法，包括以下步骤：RNA的分离纯化；探针与Marker的放射性标记和标记产物纯化；制胶和电泳；转膜与RNA分子的交联固定；预杂交和杂交；洗杂交膜和低温放射自显影；杂交探针的剥脱与膜的再利用。该方法RNA质量高，材料成本低，电泳条带整齐，噪音信号低，适合于鉴定家蚕miRNA和其他小RNA，而且对其他物种的miRNA及其靶基因鉴定也有一定参考价值。

摘要： 本发明涉及一种促进荧光原位杂交的组合物及杂交液，荧光原位杂交探针工作液、荧光原位杂交方法，属于分子病理诊断技术领域。本发明中的促进荧光原位杂交的组合物，添加了异硫氰酸胍，异硫氰酸胍能够促进探针中的DNA与样本中的DNA进行结合，提高杂交效率；因此探针工作液能够实现快速杂交、短时杂交。由该组合物制成杂交液，并调整了传统组分氯化钠、柠檬酸钠、甲酰胺、硫酸葡聚糖的浓度；得到的杂交液在3h内即可完成杂交，并且杂交后仍然具有信号亮度强并且特异性好的特点。

摘要： 本发明公开了一种针对于不同GC％含量的基因芯片杂交组件。其包括至少两种探针和杂交溶液；其中，所述芯片上探针序列GC％含量范围在10％‑80％；所述杂交溶液包含样品片段、甜菜碱和Zn2+。利用本发明基因芯片杂交组件能够使不同GC％含量的DNA片段在同一个条件下进行杂交，并可消除非特异性杂交的信号。

摘要： 本发明提供了一种简单可靠的使样品生物聚合物与探针DNA相互杂交的方法，该方法不用盖玻片和密闭容器，而是在含有与含样品生物聚合物溶液蒸气压相同的溶液的密闭容器中只使含样品生物聚合物溶液与固定有探针生物聚合物的载玻片接触，从而使样品生物聚合物与探针生物聚合物相互杂交。还提供了用于本发明的杂交微阵列和试剂盒。

摘要： 本发明涉及生物传感器技术领域，特别涉及一种荧光原位杂交芯片及荧光原位杂交方法。包括：支撑衬底和功能结构层；功能结构层设置在支撑衬底上；功能结构层具有腔体结构；腔体结构包括分离区和捕获区；分离区中设置有确定性侧向位移阵列；捕获区与分离区通过连接通道连通；捕获区中设置有捕获阵列；功能结构层中还设置有反应试剂进样通道，反应试剂进样通道与捕获区连通。通过确定性侧向位移阵列将外周血中稀少的肿瘤细胞进行富集分离，并通过捕获阵列实现了对肿瘤细胞的捕获固定。肿瘤细胞可以在捕获区中进行荧光原位杂交，从而实现了直接在荧光原位杂交芯片上进行一体化的肿瘤细胞富集及荧光原位杂交检测，简化了检测过程，省时省力。