RNA原位杂交

Materials20 ng BAC DNA clone grown o.n. in 5 mL LB + chloramphenicolAutogen reagentsSlide warmerOrganics: MeOH EtOH formamideFixative (3:1 MeOH:glacial HOAc)Biotin (or digoxigenin) nick-translation Ki ...

第四章 RNA的提取和cDNA合成第一节 概 述　　从真核生物的组织或细胞中提取mRNA通过酶促反应逆转录合成cDNA的第一链和第二链将双链cDNA和载体连接然后转化扩增 即可获得cDNA文库构建的cDNA文库可用于真核生物基因的结构、表达和调控的分析;比较cDNA和相应基因组DNA序列差异可确定内含子存在和了解转录后加工等一系列问题。总之cDNA的合成和克隆已成为当今真核分子生物学的基本手段。自 ...

IntroductionA modification of the basic immunofluorescence staining and flow cytometric analysis protocol can be used for the simultaneous analysis of surface molecules and intracellular cytokines at ...

Digoxigenin-UTP RNA labeling Digoxigenin-UTP RNA labeling 1. Add the following to a sterile microfuge tube on ice: -1 ug linearized plasmid DNA. -2 ul Dig RNA Labeling Mix 10X concentrate -2 ul 10X t ...

Digoxigenin-UTP RNA labeling Digoxigenin-UTP RNA labeling 1. Add the following to a sterile microfuge tube on ice: -1 ug linearized plasmid DNA. -2 ul Dig RNA Labeling Mix 10X concentrate -2 ul 10X t ...

Note:Start with 20 mg of total RNA for each labeling reaction.All solutions that can be filtered should be filtered.Cy dyes are light sensitive and should ALWAYS be handled in dim light.RNA Preparatio ...

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96-well RNA In Situ Hybridization Protocol The RNA in situ procedure described below is based on the protocol developed by Tautz and Pfeifle (Chromosoma 98 (1989) p81) but adapted to allow the screeni ...

Agarose-Formaldehyde ElectrophoresisRNA electrophoresis under denaturing conditions in 2.2M formaldehyde is performed according to Maniatis et al. (1982) using the MOPS buffer system. RNA under these ...

Universal RiboClone® cDNA Synthesis SystemThe Universal RiboClone® cDNA Synthesis System contains the reagents required for the synthesis of double-stranded cDNA from mRNA and subsequent ligation into ...

Universal RiboClone® cDNA Synthesis SystemThe Universal RiboClone® cDNA Synthesis System contains the reagents required for the synthesis of double-stranded cDNA from mRNA and subsequent ligation into ...

Natural History Museum Protocol for EST sequencing from lambda zap phage plaques(by PCR of phage insert cycle sequencing using ABI dye terminator kits and automated sequencing on an ABI 373A)(1) Core ...

Electrophoresis through agarose or polyacrylamide gels is the standard way to separate identify and purify nucleic acid fragments. The location of the nucleic acid within the gel can be determined ...

Electrophoresis through agarose or polyacrylamide gels is the standard way to separate identify and purify nucleic acid fragments. The location of the nucleic acid within the gel can be determined ...

In vitro RNA synthesis from plasmid-borne sequences under the control of T phage promotersN.B: Gloves should be worn at all times during preparation of in vitro RNA. All solutions should be RNase-free ...

Riboprobe® in vitro Transcription SystemsThe Riboprobe® Systems are designed for in vitro preparation of high specific activity single-stranded RNA probes or microgram quantities of defined RNA transc ...

Northern Hybridization of RNA Fractionated by Agarose-formaldehyde GelAuthor: Zuyuan QianSource: Contributed by Zuyuan QianAbstract: This is the standard and complete Northern blotting protocol includ ...

Northern Blot: Glyoxal/DMSOAdapted from Molecular Cloning (Maniatis) Current Protocols in Molecular Biology and J. Yost Lab (UMN) Objective:Glyoxal/DMSO gels are great for determining sizes of RNA esp ...

Northern BlottingAdapted from Molecular Cloning (Maniatis) Current Protocols in Molecular Biology and J. Yost Lab (UMN) Objective:Northern blotting allows detection of specific RNA sequences. RNA is f ...

Northern Blots by Michael Koelle and Tory Herman adapted from Sambrook et al. "Molecular Cloning" 4/6/94 We found that both formaldehyde and glyoxal gels work very well for electrophoresis of RNA; we ...