DNA提取与纯化

I. NEBULIZATION of DNA 1. 0.5 - 5 ug DNA in TE (10mM/1mM) 25% glycerol final volume 500 ul 2. nebulize for 90-100 sec at 5-10 psi (for a medium size of about 1.5kbp) 3. Precipitate with Et ...

BackgroundThis is the method for indirect immunofluorescence labeling; that is the antibodies do not have the fluorescent dye attached. Indirect labeling is more involved than direct labeling. If you ...

Rat liver is an ideal source for functional intact mitochondria for a number of reasons. We use Sprague-Dawley albino male rats for our studies. Female rats can be used however for serious studies we ...

AbstractGene expression and regulation are mediated by DNA sequences in most instances directly upstream to the coding sequences by recruiting transcription factors regulators and a RNA polymerase in ...

) Culture 400 mls of bacteria harboring transposon overnight at 37oC. 2) Harvest cells at 7000xg for 5 min. (pellet can be frozen at -80oC for several weeks) 3) Resuspend pellet thoroughly in 14 mls: ...

　　在真菌学研究中，传统的菌物分类以形态特征和生理生化指标为分类基础，但大部分菌物的种类多、分布广、形态特征复杂，而且少数形态特征和生理生化指标随着环境的变化而不稳定，因此，传统的菌物分类常引起分类系统的不稳定或意见分歧。近年来随着分子生物学的发展，同时也是真菌学自身发展的客观需求，分子生物学技术在真菌学研究中得到了广泛而深入地应用。一些分类地位不明确、亲缘关系不清楚的物种通过该技术得到了验证。一 ...

IntroductionNorthern blotting nuclease protection assays and RT-PCR are frequently used to analyze gene expression. While qualitative results from these assays are sometimes sufficient most researcher ...

一、什么是克隆技术 　　克隆，是Clone 的译音，意为无性繁殖。 英语"Clone"一词起源于希腊文"Klone"，原意是用"嫩枝"或"插条"繁殖。 现在"克隆"的含义已不仅仅是简单的"无性繁殖"，凡来自一个祖先，经过无性繁殖出的一群个体，也叫"克隆"。这种来自一个祖先的无性繁殖的后代群体也叫"无性繁殖系"，简称无性系。在自然界，有不少植物具有先天的克隆本能，如番薯、马铃薯、玫瑰等插枝繁殖的植物 ...

做分子生物学实验，核酸纯化、酶切、克隆算是最基本的步骤了。因为要做定向克隆，通常要作双酶切。为了省时省事儿，我们常常想方设法把两个酶放在一起切。可是事情并非总那么称心如意――两个酶经常在一起闹别扭，不是反应温度不同啦，就是Buffer不同，有时候两个酶切位点连在一起，必须先切一个再切另一个，否则就切不动――因为有的酶除了识别位点之外还需要旁边留有几个碱基，如果正好被切掉了就出问题。于是有人 ...

Note: The pointers in this technical bulletin presuppose the use of standard 50% formamide hybridization solutions. However if ULTRAhyb™ Ambion's ultrasensitive hybridization buffer is used the differ ...

Northern assays require RNA (total- or poly(A)- selected) to be resolved on a agarose gel under denaturing conditions transferred to a membrane and immobilized for subsequent hybridization. Various in ...

Analysis of mRNA expression in tissue or cell culture is often done by Northern blot or ribonuclease protection assay (RPA). Northern assays require the total RNA to be resolved on a denaturing agaros ...

1. Cut 10ug of plasmid DNA where you want the transcript to end.2. Add an equal volume of 2X PK buffer:200mM TrisHCl pH 7.525mM EDTA300mM NaCl2% SDS200ug/ml proteinase K3. 37� for 30min and then extra ...

Microarray analysis and differential display have become popular techniques for identifying differentially expressed genes. Once identified the varying expression levels of specific mRNAs must be conf ...

1. Add appropriate volumes of RNA ( Try 5 and 10 ug RNA plus 600 or 800 pg dig labelled probe.2. Include 2-3 yeast RNA samples/probe used. 2ul of 5mg/ml stock.3. Add 20 ul of Soln. A to all of the tub ...

RNase Protection Assay - ProtocolThis method can be used to detect and quantitate mRNA to map mRNA termini and to determine the position of introns within the corresponding gene. High specific activit ...

End-label oligo (20-25 mer)4 µl 5X T4 Kinase Buffer5 µl ATP (7000 Ci/mmol)10 µl water + oligo (200 ng)1 µl T4 Kinase1. 37 deg C for 1 hour.2. Heat inactivate at 75 deg C for 10 minutes.3. Add 1 µl gl ...

全文下载：http://www.ebioe.com/down/html/down_211.htm陈秀英 彭孝军~A大连理工大学精细化工国家重点实验室 大连 116012)摘 要 介绍了各种DNA 荧光探针的结构特征、荧光性质和与DNA 的作用方式，概述了DNA 探针在生物分子分析方面的应用，并展望了DNA 荧光探针的发展趋势和应用前景。关键词 荧光探针 DNA 花菁染料 光稳定性 量子点DNA F ...

体外定点突变技术是研究蛋白质结构和功能之间的复杂关系的有力工具，也是我们在实验室中改造/优化基因常用的手段。蛋白质的结构决定其功能，二者之间的关系是蛋白质组研究的重点之一。对某个已知基因的特定碱基进行定点改变、缺失或 者插入，可以改变对应的氨基酸序列和蛋白质结构，对突变基因的表达产物进行研究有助于我们了解蛋白质结构和功能的关系，探讨蛋白质的结构/结构域。而利用定点突变技术改造基因，相信大家也非常熟 ...

一、 目的掌握质粒的小量快速提取法；了解质粒酶切鉴定原理。二、 原理质粒（plasmid）是一种染色体外的稳定遗传因子。大小在1~200kb 之间，具有双链闭合环状结构的DNA 分子。主要发现于细菌、放线菌和真菌细胞中。质粒具有自主复制和转录能力，能使子代细胞保持它们恒定的拷贝数，可表达它携带的遗传信息。他可独立游离在细胞质内，也可整合到细菌染色体中，它离开宿主的细胞就不能存活，而它控制的许多生物 ...