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现代分子生物学研究发现，中药材(不含矿物药)所依赖的生物资源――“物种”的多样性是由于其基因多态性的结果，而基因多态性可在分子水平上检测，它比在形态、组织和化学水平上检测更能代表其变异类型的遗传标记。传统的分子标记主要有血清学特征、蛋白质谱 、同工酶谱，近10年来则有迅速发展起来的DNA 分子标记，现介绍如下。 1 　传统的分子标记 1.1　抗血清　中药有许多具有抗原决定 ...

Pyrosequencing是对短到中等长度的DNA序列样品进行高通量的、精确和重复性好的分析的技术。 第一步——测序引物和PCR扩增的、单链的DNA模板杂交，与酶—DNA聚合酶（DNA polymerase）、ATP硫酸化酶（ATP sulfurylase）、荧光素酶（luciferase）、三磷酸腺苷双磷酸酶（apyrase）—和底物&mdash ...

1.对于微量DNA样品溶液（如100ml以下），应预先将DNA溶液以TE或无菌水稀释至一定体积。以减少第5步中的DNA损失。 2.加入等体积的酚:氯仿:异戊醇（25:24:1）。 3.剧烈振荡管中内容物，将水相和有机相充分混匀，使之成为乳浊液。 4.室温下静置以初步分层。再用离心机于室温以10000rpm离心至少5分钟以上，使无机相与有机相充分分开。 5.用剪去尖头的微量移液器枪头小心移出 ...

本法的产量要低得多，但却比本章所述的其他方法更温和，是提取大质粒(大于15kb)的首选方法。 试验步骤: 1、将500ml细菌沉淀物洗过后重悬于10ml用冰预冷的10%蔗糖、50mmol/L Tris.Cl(pH8.0)溶液中，将悬液移至30ml的带盖螺口塑料试管中。 2、加入2ml新配制的溶菌酶溶液。当溶液的pH值小于8.0时，溶菌酶不能有效地发挥作用。 3、加8ml 0.25mol/L EDT ...

Materials: • 0.8 % agarose gel in 1x TAE • Digested DNA • Glass Milk • NaI solution • New Wash Procedure: 1)Run digested DNA out on agarose gel slowly (70 V on BioRad gel) 2)U ...

1.Grow 5 ( large scale-15ml culture). Harvest in single eppendorf tube (or 15 ml disposable tube). 2.Resuspend pellet with 300μl STET buffer (900μl). After resuspending add 30μl RNase/lysozy ...

Hancock Laboratory Methods. Department of Microbiology and Immunology University of British Columbia British Columbia Canada http://www.cmdr.ubc.ca/bobh/showmethod.php?methodid=12 REFERENCE: GATA 19 ...

Amberg Lab Upstate Medical University http://www.upstate.edu/biochem/amberg/protocols/seq_temp.html Notes on DNA : The cleaner the better.If mini DNA use phenol extracted and use the entire mini pre ...

Section of Cancer GenomicsGenetics BranchNCI National Institutes of Health Reagents Chloroform EDTA0.5 M Ethanolabsolute Isoamyl alcohol SigmaCat.I-3643 Phenol Phosphate Buffered Saline (PBS)1X Protei ...

Cepko/Tabin Lab Harvard University http://axon.med.harvard.edu/~cepko/protocol/mike/E1.html Cepko/Tabin Lab Harvard University http://axon.med.harvard.edu/~cepko/protocol/mike/E1.html Solutions 10X A ...

when I extracted DNA from chicken red blood cellI met a very strange question.After the phenol and chloroform treation.I collected the upper layers (it is not liquid but like some transparent s ...

The Minion Lab College of Veterinary Medicine at Iowa State University http://mycoplasmas.vm.iastate.edu/lab_site/methods/DNA /restrdig.html ...

Overview Procedure 1. Preheat the CTAB Isolation Buffer at 60℃. 2. Grind 2 g of fresh leaf tissue to a powder in Liquid Nitrogen in a chilled mortar and pestle. 3. Scrape the powder into a chilled 50 ...

Prepare: 200ml LB in a 1L flask 1L sterile ddH2O and place in cold room 4 50ml conicals chilled on ice 10% glycerol (cold) Chilled boxes of 100μl and 1ml pipet tips. Chilled 0.5ml tubes. General Th ...

RNA/Zeta Probe Dot Blotting protocol 1)Make up RNA (up to 20μg)dissolved in sterile H2 OTE or 0.5% SDS to 500μl with ice-cold sterile 10mM NaOH1mM EDTA and apply it to Zeta Probe membraneheld in ...

长时间反应时内切酶的活性保持情况因酶而异。本表中列出了在16小时内完全酶解底物DNA所需的最小酶量。 实验方法：在50µl的反应体系中，分别加入1µg的单位定义底物DNA和1.00、0.50、0.25和0.13个单位的内切酶，37℃（或要求的最适温度）反应16小时。用琼脂糖凝胶电泳法来确定在16小时内酶解1µg底物DNA所需的最小酶量。 16小时酶 ...

Retroelements and their derivatives are a ubiquitous and abundant component of plant genomes. From the 1990s PCR based techniques have been developed to isolate the elements from genomic DNA of differ ...

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 ...

The following protocol is based on our modifications of R. Kraft J. Tardiff K. S. Krauter and L. A. Leinwand. Biotechn . 6(6):544-545 1988. 1.Inoculate 2-5 ml of L broth containing the appropriate ant ...

1.Wash the plates briefly using the 96-channel block washer. 2.Place the plates in a tub submerged in dd-water. 3.Put a flask on top of the plates to keep them submerged. 4.Place the tub with the s ...