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质粒提取简介及问题分析
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摘要:一、导论 质粒提取的原理:转自复旦大学一位老师的帖子,后面是方法介绍 碱裂解法从大肠杆菌制备质粒,是从事分子生物学研究的实验室每天都要用的常规技术。可是我收研究生十几年了,几乎毫无例外的是我那些给人感觉什么都[阅读全文:]

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Labeling oligonucleotides with 32P ATP
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摘要:Wear gloves throughout and work in radiation area. Monitor area before and after use.Mix the following in an eppendorf tube:1. 0.5 microgram oligonucleotide dissolved in H2O.2. 3 microliters 10x kinas[阅读全文:]

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Removal?of?32P-ATP?from?Oligonucleoi
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摘要:1. Prepare Whatman DE-52 according to manufacturers specifications. Equilibrate at store with 0.02% Sodium azide.2. Plug a 1 mL (blue) pipet tip with Siliconized glass wool. Add 500 ml of 50:50 DE-52 [阅读全文:]

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Salmon Sperm DNA (10mg/ml)
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摘要: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) [阅读全文:]

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DNA Recovery With Low Melt Agarose
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摘要:Recovery of DNA from Low Melting Point Agarose Gels1.Run digestion products on 0.7% LMP agarose gel in 1X TBE (it's nice to have at least 1ug of the fragment you want). LMP agarose is fragile; pour ge[阅读全文:]

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酵母菌基因组DNA的提取
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摘要:酵母 菌基因组 DNA的提取一:仪器: 同方法一二:试剂: SE缓冲液(1M山梨醇,0.1MEDTA pH7.5);溶菌酶(50mg/ml);20%PVP;蛋白酶K缓冲液(10mM Tris pH7.60.5% SDS 1mM EDTA);其余同前三:操作1.5ml对数生长期细菌细胞离心,120[阅读全文:]

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UV?Quantitation?of?DNA
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摘要: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 n[阅读全文:]

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Improved?Alcohol?Precipitation?of?DNA
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摘要: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 fin[阅读全文:]

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Deproteination?using?phenol/chloroform
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摘要:'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. Th[阅读全文:]

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RLGS?protocol
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摘要: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[阅读全文:]

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DNA技术