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plasmid

[核酸技术] 2008年05月份

Mini

标签:DNAplasmidandMiniExtractionIntroductionPurification

Introduction Extraction and purification of plasmid DNA from E.coli can be performed on a small scale,where DNA is extracted from just 1-2 ml of bacterial culture (mini-prep), or from much larger volumes (100 ml maxi-prep; 1 L giga-prep). The procedure re

标签:plasmidandPurposePurificationShuttle

1.0 Purpose 1.1 This protocol may be used for the linearization and purification of the shuttle plasmid. 2.0 Materials 2.1 Equipment DNA Gel Electrophoresis Apparatus Heating block set at 65 C Centrifuge -20 C Freezer UV Spec 2.2 Supplies Restriction Enzy

标签:DNA分离plasmidIsolation碱裂解法AlkalineLysis

Reagents: Soln 1: 50 mM glucose/10 mM EDTA/25 mM Tris pH 8. Autoclave before use. Add 2 mg/ml Lysozyme just prior to use. Soln 2: 0.2 N NaOH/1% SDS. Keep solution at room temperature. Solution is normally prepared from 10X concentrated solutions the day o

标签:DNAPEGplasmid制备乙二醇Preparation

Plasmid DNA isolated by this procedure can be used routinely for electrophoretic analysis, restriction endonuclease digestion and transformation of E. Coli., DNA sequencing, PCR and most other molecular biological techniques. The procedure is a modificati

标签:DNAplasmid制备Prep碱裂解法AlkalineLysis

Alkaline lysis miniprep 1. Grow bacteria overnight in 37° shaking incubator, with lids very loose and taped on. I normally use 5 ml of liquid medium in a 50ml conical bottom tube. Be sure to include the appropriate selective antibiotic. 2. Remove 1 ml of

标签:plasmidMiniNotesbeforeprotocolPurificationImportant

Important Notes Before Starting New users are strongly recommended to read the QIAGEN handbook before starting the procedure. Before using the kit for the first time, dissolve the lyophilized RNAse A provided Buffer PI . Buffer PI should then be stored at

标签:纯化plasmidMiniThisprotocolPurification

This protocol is for Mini (up to 20 g) preparations of high-copy plasmid DNA from cultures of E. coli . Important notes before starting New users are strongly recommended to read Appendix A (page 21) at the end of this handbook before starting the procedu

标签:plasmidKitProcedureGenEluteTMMiniprep

Note All centrifugation speeds are given in units of g. Please refer to Table 1 for information on vonverting g -force to rpm. If centrifuges/rotors for the required g -forces are not available, use the maximum g -force possible and increase the spin time

标签:携带plasmid制备外源

质粒是携带外源基因进入细菌中扩增或表达的主要载体,它在基因操作中具有重要作用。质粒的分离与提取是最常用、最基本的实验技术。 质粒的提取方法很多,大多包括3个主要步骤:细菌的培养、细菌的收集和裂解、质粒DNA的分离和纯化。本实验以碱裂解法为例,介绍质粒的抽提过程。 实验目的 :掌握碱裂解法抽提质粒的原理、步骤及各试剂的作用。 实验材料 :含有质粒

标签:DNAplasmid制备微量

实验原理 质粒(Plasmid)是一种双链的共价闭环状的DNA分子,它是染色体外能够稳定遗传的因子。质粒具有复制和控制机构,能够在细胞质中独立自主的进行自身复制,并使子代细胞保持它们恒定的拷贝数。目前,质粒已广泛的用作基因工程中目的基因地运载工具—载体。从大肠杆菌中提取质粒DNA,是一种分子生物学最基本的方法。 质粒DNA分离纯化方法有多种,但其原理和步骤

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