DNA提取与纯化

macerate tissue in Eppendorf tube without butter at RT add 400 m l extraction buffer vortex for 4 sec leave sample at RT until other samples are ready ( 1 h) spin in microfuge for 1 min transfer 300 m ...

PEG Preparation of Plasmid Plasmid isolated by this procedure can be used routinely for electrophoretic analysis restriction endonuclease digestion and transformation of E. Coli. sequencing PCR and mo ...

ECKDescriptionThis method was suggested in a BRL "Focus" article several years ago (Zeugin & Hartley 1985). It is a useful way to avoid coprecipitation of proteins and accumulation of salt in the fina ...

The principle and procedures of RLGS method was first described by Hatada et al. (1991) and its improvement was described by Asakawa (1996). Basically based on their procedures we have successfully ap ...

DNA absorbs ultraviolet light due to its highly conjugated nature. DNA may thus be easily quantitated in a UV spectrometer.Typically 1 OD260 (i.e. a solution having an absorbance of one unit at 260 nm ...

Sample DNAs should not be highly degraded nor contain PCR inhibitors such as high concentrations of heme or chelating agents. For each individual assayed 250 ng of genomic DNA are digested separately ...

'Phenol' as used is Analar grade. Phenol should be melted at 65℃8-hydroxyquinoline added to a final concentration of 0.1% and equilibrated three times with an equal volume of 1M Tris.HCl pH 7.0. The f ...

Salmon Sperm DNA1.Dissolve 1 g salmon sperm DNA in 100 ml H2 O. 2.Autoclave (20 minutes) and aliquot in 1 ml/tube 3.Store at -20℃. (common freezer for stock solutions) ...

用于Genome Walking Kit的模板总DNA 一般要满足以下几点：1、DNA 的完整性，电泳为单一条带。2、避免杂质，OD260/280=1.8~2.0。3、模板DNA 不要被污染。4、准确定量，不要少于3μg。 ...

Protocols for HapMap assay-design Affymetrix platform (used by Broad)Defined protocols:LSID: urn:LSID:affymetrix.hapmap.org:Protocol:affy_assay_design_1:1Title: Genotyping using Affymetrix arraysDescr ...

FINGERPRINTING PROTOCOL (AGAROSE GEL) * Restriction digests consist of:15.75 ml ddH2 O 1 µl 10 X buffer B (Boehringer Mannheim) 0.25 µl HindIII (40 U/µl) 3 µl DNA Set up digestions in 96 well plates. ...

【实验目的】掌握植物总DNA的抽提方法和基本原理。学习根据不同的植物和实验要求设计和改良植物总DNA抽提方法。【实验原理】通常采用机械研磨的方法破碎植物的组织和细胞，由于植物细胞匀浆含有多种酶类(尤其是氧化酶类)对DNA的抽提产生不利的影响，在抽提缓冲液中需加入抗氧化剂或强还原剂(如巯基乙醇)以降低这些酶类的活性。在液氮中研磨，材料易于破碎，并减少研磨过程中各种酶类的作用。十二烷基肌酸钠(sark ...

Q:my group is involved in a national project to develop a genomic platform for cancer research. while Nimblegen has developed target enrichment chip for the Genome Sequencer 20 nothing similar has bee ...

【实验目的】（1）学习并掌握动物组织总DNA的提取方法及其原理。（2）从肝脏组织中提取到一定量的纯净的DNA样品。【实验原理】DNA是一切生物细胞的重要组成成分，主要存在于细胞核中。通过研磨和SDS作用破碎细胞；苯酚和氯仿可使蛋白质变性，用其混合液（酚：氯仿：异戊醇）重复抽提，使蛋白质变性，然后离心除去变性蛋白质；RNase降解RNA，从而得到纯净的DNA分子。【仪器、材料、试剂】（一）仪器1．高 ...

按照RNA分离操作方案在完全移去水样层后，匀浆中的DNA存在于中间层和苯酚层中也可以被分离出来。在沉淀和多次洗脱后，DNA溶解在8 mM NaOH中。用TRIZOL试剂从组织和培养细胞中完全回收的DNA可以用来做样品中DNA含量的测定。同时抽提的基因组DNA可以用于对Northern analysis的结果进行标准化，因为DNA变异程度比总RNA或组织重量要小。实验所需但试剂未提供的物品：_酒精 ...

2. Materials2.1. MCARestriction enzymes SmaI XmaIT4 DNA ligaseTaq DNA polymerase10X PCR reaction buffer:670mM Tris-HCl pH 8.840mM MgCl2160 mM NH4(SO4)2100 mM b -Mercaptoethanol1 mg/ml bovine serum alb ...

Hi I am a Biochem student. �I am extracting DNA from plants that have been exposed to contaminants then using restriction enzymes to see if there is any change. �I was wondering if there was a differe ...

Fungal Genomic DNA ExtractionOverviewHigh throughput of many fungal isolates can be achieved by growing axenic cultures in either (a) 1.5mL microfuge tubes half full with liquid media (500uL) with a h ...

Tissue collection storage microdissection sectioning: See separate protocol. Tissue handling: Note that all fresh tissue should be handled as BioSafety Level 2 materials (wear gloves lab coat etc.). D ...

一、直接检出病毒核酸 　　1．核酸杂交（Nucleic acid hybridization） -临床病毒学中报速诊断方法通常是检测标本中的病毒抗原，然而核酸分子杂交具有高度敏感性和特异性，斑点杂交 (Dot hybridization) 广泛用于检测呼吸道标本，尿标本中的病毒核酸。标本滴加到硝酸纤维素膜上，病毒DNA结合到膜上，在原位进行硷变性处理后，有放射标记的已知病毒DNA片段杂 ...