DNA实验技术

Hahn Lab，The Fred Hutchinson Cancer Research Center and Howard Hughes Medical Institute http://www.fhcrc.org/science/labs/hahn/methods/genetic_meth/yeast_transf_rapid.html ...

一、目的： 了解分离提取DNA的一般原理，掌握从动物肝脏中提取DNA的方法。 二、原理： 在浓氯化钠（1—2mol·L-1）溶液中，脱氧核糖核蛋白的溶解度很大，核糖核蛋白的溶解度很小。在稀氯化钠（0.14mol·L-1 ）溶液中，脱氧核糖核蛋白的溶解度很小，核糖核蛋白的溶解度很大。因此，可利用不同浓度的氯化钠溶液，将脱氧核糖核蛋白和核糖核蛋白从样品中分别抽提 ...

Uniplex PCR-based Sequencing Latest update 2-21-00 A.PCR Reactions: B.Processing PCR products: C.Alternative PCR Reaction Cleanup steps instead of passage through Sephadex G-50 usually not performe ...

影响因素 1.肿瘤标志物具有特异性和非特异性的双重特点。 2.各种肿瘤特性的不同，如肿瘤的分期、大小、定位、组织来源及转移趋向等的不同。 3.各种抗体的不同，如多克隆抗体、单克隆抗体、抗体决定簇和抗体亲和性等的不同。 4.各种检测系统的不同，如检测原理、标记物、实验条件、检测技术、检测仪器和检测敏感度等的不同。 5.检验人员的专业水平和技术经验、以及实验室条件和实验的可靠性等的差异。 ...

When just a few transformants are sufficient such as the transformation of a test plasmid or a GAL4BD fusion plasmid the following protocol can be used. This procedure is very flexible and can be appl ...

1. Pour a vertical acrylamide gel using TEA buffer. A 4 % non denaturing gel is correct for most applications. 2. Run out DNA fragments. For fragments greater than 500 bp run the xylene cyanol dye to ...

Procedure for 100 ~ 300 mg of tissue. 1. Collect fresh tissue coleoptiles or small amount of leaf tissue. Use about three inches of a leaf blade. Preferably younger tissue. Fold leaf and place i ...

I am working with birds of prey....i am looking for a simple method for isolating genomic DNA from avian breast muscle....I have found protocols for mammalian tissue...will these work fine? als ...

DNA Extraction from Archival Formalin-fixed Paraffin-embedded Tissue Sections Author: Shi et al. Source: Contributed by APostodoc Abstract: Describes two methods of extracting DNA from archived paraff ...

DNA是通过异羟基洋地黄毒苷（digoxigenin，Dig）配基标记的脱氧尿嘧啶核苷三磷酸（dUTP）随机插入结合而被标记。dUTP通过间臂连结类固醇半抗原异羟基洋地黄毒苷酸基，形成Dig-dUTP，杂交反应后，杂交的靶DNA通过酶联免疫法与一个抗体复合物结合，接着在5-溴-4氯-3-吲哚磷酸盐（X-磷酸盐）和硝基四氮唑蓝（NBT）存在下，由酶催化反应，在杂交部位形成蓝紫色带或颗粒 ...

(adapted from Bruce A. Roe Department of Chemistry and Biochemistry The University of Oklahoma Norman Oklahoma 73019 broe@ou.edu) Typically 2.5 - 3 volumes of an ethanol/acetate solution is added to t ...

TaKaRa的内切酶和NEB的内切酶哪个更好一些？ 参考见解： TaKaRa的内切酶、NEB的内切酶两个公司的酶的品质都非常好。NEB公司的酶的活性很高，切出两条小带可能是因为出现星活性，可以试试把酶量减半。NEB的酶很多都是克隆的，所以纯度比较高。 应用磁性微球提取人全血基因组DNA，将提取出的DNA直接用于限制性酶切反应，应用Taq1酶，但发现酶切后产生的是smier片断，即切碎的状态。不知 ...

Has anyone any experience of using a semi dry blotter for Southern blotting. At present I am tranfering overnight using a weight I think there is a blotter that is suitable for nucleic acids and can t ...

Growth and storage of Agrobacterium tumefaciens Strain GV3101: resistant to gentamycin and rifampicin so add 25-50 ug/ml Gentamycin 10 ug/ ml rifampicin on plates or in liquid media for selection. GV3 ...

Preparation of electroporation cells 1. Prepare an overnight of NM522 in minimal medium. 2. Inoculate 1L LB with 10ml (1/100th vol) of the overnight and grow to A600 = 0.5- 1.0. 3. Pellet cells 5krpm ...

碱法提取的质粒DNA即使用RNA酶处理，仍会含有少量RNA。当有些试验需无RNA污染的DNA制品时，则需进行进一步纯化。一般常用Sepharose 2B或Sepharose 4B进行纯化，该方法具有快速，条件温和，重复性好，载体物质可以再利用等优点，因而已广泛用于质粒DNA纯化。 1、将Sepharose 2B经含0.1% SDS的TE（pH8.0）平衡后上柱。 2、将至多1ml的DNA溶液铺 ...

在已知基因部分核苷酸序列或全部核苷酸序列的情况下，基因的制备方法主要可分为两大类：一类是基因化学合成法，一类是基因生物制备法。一般又可将基因生物制备法分为文库法、cDNA 文库法和PCR法等。 1 化学合成法。 就化学本质而言，基因是一段具有特定生物功能的核苷酸序列。如果知道了基因的分子结构，就可以进行基因的化学合成。有关DNA 的化学合成方法主要有磷酸二酯法、磷酸三酯法、亚磷酸三酯法、亚磷酸酰胺 ...

The Preuss LabThe Division of Biological SciencesThe University of Chicago. http://preuss.bsd.uchicago.edu/protocols/enzyme.html ...

Acid Washing Beads Peter Novick Lab Department of Cell Biology Yale University School of Medicine http://info.med.yale.edu/cellbio/Novick/Second/Protocols/AcidBeads.pdf Materials: 1) 0.5 mm glass bead ...

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 norm ...