DNA实验技术

・ RNA Injection (Hoshi Lab)The oocyte is a useful model for the investigation of the function of usr/localious genes and is a widely used system for the translation of microinjected natural an ...

・ Designing Your Peptide (Genosys) ・ Handling & Storage of Peptides (Genosys) Two Dimensional Peptide Mapping (Sefton Lab) Synthesizing Peptides (Perkin-Elmer) Solid Phase Peptide ...

HeLa Cell Nuclei Preparation (John Garland)Prepare nuclear extract from HeLa cell ・ Extract Preparation (Brent Graveley)Preparation of nuclear extract and cytoplasmic extract ・ Pol ...

・ Protein detection (Aberdeen's Lab)The method used to locate the proteins following 2D-PAGE depends on the nature of the original sample.・ Protein detection(Silver staining) (ExPASy)T ...

Western 杂交（主要内容如下）Preparing of Protein Lysates Western Blotting Far Western Blotting Semi Dry Blotting Stripping Membranes Trouble Shooting and Others Preparing Protein LysatesPreparing Protein Lysate ...

蛋白质提取和纯化（主要内容如下）Protein Extraction Protein Purification Protein Precipitation Column Preparatioin Q & A Posted in the Method Forum Protein Extraction Whole Cell Lysate Preparation (UCSF Tumor Immunol ...

Biosynthetic labeling (Sefton Lab) Biotinylation of Antibody (Contributed by Nanci Donacki) 125I Labeling of Protein using ICl (ScienceXchange) Protein (Antibody) Fluorescent Labeling (Mario Roederer) ...

Protein Construct Expression and Purification Procedures(Gimila's Lab) Protein Expression (Mark's Lab) ・ Purification of GST Fused Proteins (Contributed by Chia Jin Ngee)Many people have ...

Interaction Trap/Trap Two-Hybrid System ・ Yeast Two-Hybrid System (Finley Lab)This is one of the most comprehensive and detailed guide to yeast two-hybrid system technique with introduction to ...

Tyrosine Kinase Assay Using Synthetic Peptides (T. Miller)Small synthetic peptide substrates are especially well suited for applications such as assays of tyrosine kinases in permeabilized cells or fo ...

・ In Vitro Translation (Promega)Provides general protocol for coupled single-tube tscription/translation reactions for eukaryotic in vitro translation. ・ In vitro Protein Synthesis (P ...

Serial Analysis of Gene Expression (SAGE)SAGE is a powerful tool that allows the analysis of overall gene expression patterns with digital analysis. Because SAGE does not require a preexisting clone ...

Randomly Amplified Polymorphic DNA (RAPD) Randomly Amplified Polymorphic DNA (RAPD) by (DNA KAFFE)RAPD analysis has been successfully used in mapping and fingerprinting applications. RAPD can efficie ...

Serial Analysis of Gene Expression (SAGE)SAGE is a powerful tool that allows the analysis of overall gene expression patterns with digital analysis. Because SAGE does not require a preexisting clone ...

DNA Fingerprinting (David F. Betsch)the theory procedures and applications ・ DNA Fingerprinting (Polyarylamide Gel) (Caltech)BAC DNA sample and gel preparation gel running and more ・ ...

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 ...

核糖核酸探针1．室温下，于1.5ml无菌微量离心管内依次加入：5μl 5×转录缓冲液1μg 模板DNA1.2μl 10 mmol/L rATP（终浓度为480μmol/L）1.2μl 10 mmol/L rCTP（终浓度为480μmol/L）1.2μl&n

放射性标记的探针与固定在膜上的核酸Southern杂交1. 将含有靶DNA的膜漂浮在盛有6×SSC（或6×SSPE）的皿中直到膜自下面而上完全浸湿。将膜浸于溶液中2 min。2. 用下列方法之一进行预杂交在热密封的袋中进行的杂交a. 将膜塞入热密封袋中（如Sears Seal-A-Meal或相应设备），按每平方厘米0.2ml加入欲杂交液中。尽量将袋中的空气挤出。b. 用热封口 ...

用于检测植物组织RNA的原位杂交法（一）组织切片制备l 植物组织的甲醛固定和包埋1．将植物组织切成小块，立即放入一个装有10～50ml固定剂溶液的烧杯中，把烧杯放到真空脱水机内。调节真空度以形成一个温和的真空，然后慢慢恢复常压。微量便于固定剂渗入组织，必须反复真空和非真空状态。待固定剂充分渗入组织块后，室温下放置2～3小时。2．室温下用50～100ml 50mmol/L PIPES缓冲液 ...

确定各种可能与目标蛋白相互作用的蛋白质，“撒大网”l 双杂交和其他双成分系统第一阶段：诱饵-LexA融合蛋白的鉴定诱饵-LexA融合蛋白的构建1．将编码诱饵蛋白的靶DNA克隆到LexA融合载体的多聚接头处，以合成一种框架内的LexA融合基因。确定诱饵序列的羧基端存在翻译终止序列。形成的质粒作为pBait。2．采用下列LexA融合基因和lexAop-lacZ报道质粒的组合，建立一系列E ...