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DNA制备

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DNA标记

DNA标记(主要内容如下) DNA Labeling by Nick Translation Random Primed Labeling End-Labeling Purification of Labeled DNA Non-isotopic Labeling Others DNA Labeling by Nick Translation§ DNA Lab ...

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DNA克隆

DNA克隆(主要内容如下)・ General Procedure ・ PCR Cloning ・ Subcloning ・ ET Cloning ・ Vector Preparation ・ Ligation Reaction ・ Colony Screening General Pro ...

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DNA测序

DNA测序(主要内容如下)・ Sequencing Gel Preparation ・ Preparation of Templates ・ DNA Sequencing by the Dideoxy Method ・ DNA Sequencing by the Chemical Method ・ Dye Termi ...

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RNA标记

RNA labeling (Amersham Pharmacia)For the generation of radiolabelled single stranded RNA for use as hybridization probes 32P-pCp 3' End-labeling RNA (Jim Brown Lab)This procedure includes two metho ...

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RNA电泳

・ RNA Gel (Crawford Lab)・ Gel Electrophoresis of RNA (Beverly Faulkner-Jones)great tips on RNA gel electrophoresis.・ Northern Gel and Transfer (gopher://iubio.bio.indiana.edu ...

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cDNA

・ cDNA Synthesis (Crawford Lab)mRNA can be converted into DNA (copy DNA cDNA) by annealing oligo-dT to the 3' poly-A tail that occurs on all eukaryotic mRNA. After the dTs bind to the As we wi ...

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引物延伸反应

Primer extension analysis is used to determine the location and quantitate the amount of the 5´-end of specific RNAs. An end-labeled oligonucleotide is hybridized to RNA and is utilized as a primer b ...

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mRNA末端快速扩增

mRNA末端快速扩增(RACE)Rapid Amplification of mRNA Ends by PCR (RACE) (PMCI Research) ...

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蛋白质定量

Quantitative Determination of Peptides by Sulfhydryl (-SH) Groups New (Contributed by David Van Horn Dept. of Chemistry UC Berkeley Greg Bulaj Dept. of Biology University of Utah)This is the standard ...

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蛋白质电泳

蛋白质电泳(主要内容如下)One-Dimensional SDS-PAGE Two-Demensional SDS-PAGE Protein Electrophoresis in Agarose Gel Gel Staining Recipes One-Dimensional SDS-PAGE ・ Protein Gel and Staining (Gottschling Lab ...

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遗传作图

Gene Mapping (Amersham)DNA extraction PCR amplification and gel resolution. ...

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基因治疗

A Simplified System for Rapid Generation of Recombinant AdenovirusesGive detailed guide to the construction of recombinant adenoviruses. ...

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足迹法

High Resolution Genetic Footprinting (Stanford)Genetic footprinting is a technique for high-resolution mapping of the functional organization of a cloned gene. An in vitro transposition reaction with ...

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酵母转化

・ Yeast Transformation (Gietz Lab)LiAc/SS-DNA/PEG Transformation・ Yeast Transformation (Breeden Lab)LiAc method・ Large-Scale Yeast Transformation (Yale Yeast Analysis Center)De ...

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酵母培养基和相关试剂

Yeast Hartwell's Complete (HC) Media (Gottschling Lab) Yeast Complete Media Yeast Media (PMCI) Yeast Media Solutions and Stocks (Donis Keller Lab) HC Dropout Plate Supplementation Table (Gott ...

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酵母准备

Yeast DNA Preparation Yeast Genomic Preparation (Gottschling Lab)Rapid method for yeast genomic DNA isolation Yeast DNA Preparation (rapid glass bead prep) (Hahn Lab) Yeast Genomic DNA Preparatio ...

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利用聚合酶链反应制备放射性标记的DNA探针

利用聚合酶链反应制备放射性标记的DNA探针1. 在一个0.5 ml的薄壁微量离心管中设置下列扩增/放射性标记反应体系:10×扩增缓冲液 5.0μl10 mmol/L dNTP溶液 1.0μl0.1 mmol/L dCTP 1.0μl20μmol/L 正向寡核苷酸引物 2.5μl20μM反向寡核苷酸引物 ...

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T4多核苷酸激酶末端标记

用T4标记的5’末端1.无菌的1.5ml为了离心管置冰浴上顺序混合下列组分:50 pmol合成的寡核苷酸2.5μl 10×反应缓冲液10μl γ-PATP(33pmol)1.5μl T4多核苷酸激酶(15U)灭菌双蒸水加至25μl2.37℃温育30min。3.当反应物正温育时,以2000×g离心2min制备Sephadex G-25自旋柱。4.加入2μl 0.5mol/L EDTA或加热至68℃ ...

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应用激酶交换反应的末端标记

应用激酶交换反应的末端标记1.在置冰浴中的1.5ml微量离心管内依次加入:50 pmol合成寡核苷酸2.5μl 10×反应缓冲液1.5μl ADP(300μmol/L终浓度)10μl γ-PATP(33pmol)1.5μl T4多核苷酸激酶(15U)蒸馏水加至25μl2.37℃水浴温育30分钟。3.加入2μl 0.5mol/L EDTA或加热至68℃ 10min终止反应。混合液经Sephadex ...

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Northern杂交

Northern杂交1.在10~20 ml 预杂交液中68℃温育膜2h。2.若使用双链探针,在100℃下加热32P标记的双链DNA 5 min,使之变性,然后迅速移至冰水中冷却。3.把变性的或单链放射性标记的探针直接加到预杂交液中,在合适的温度下继续温育12~16h。4.杂交后,将膜从塑料袋中取出,在室温下迅速转移到含有100~200ml 1×SSC,0.1%SDS的塑料盒内,将盒盖好,置于水平振 ...

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