DNA实验技术

pGL3 Luciferase Reporter VectorsThe pGL3 Luciferase Reporter Vectors provide a basis for the quantitative analysis of factors that potentially regulate mammalian gene expression. These factors may be ...

pGL3 Luciferase Reporter VectorsThe pGL3 Luciferase Reporter Vectors provide a basis for the quantitative analysis of factors that potentially regulate mammalian gene expression. These factors may be ...

TRANSFORMATION (ONE-STEP PEG METHOD)Dilute 1:100 a fresh O/N culture of bacteria into prewarmed LB broth and incubate cells with shaking (225 rpm) to an OD600 of 0.3-0.4. Add an equal volume of ice-co ...

Transformation of Competent CellsA. Preparing competent cells.Inoculate 30 ml of SOB broth in a 250-ml flask with bacteria to be transformed from a single colony on a fresh plate. Incubate overnight a ...

Transformation of E. coliReference: H. Inoue H. Nojima & H. Okayama Gene 96 23-28 (1990) 1. Culture E. coli in 250 ml of SOB medium in a 2-liter flask at 18 C with vigorous shaking until an A260 of 0. ...

Bacterial Colony PCRObjective:This protocol allows rapid detection of transformation success when primers are available to allow determination of correct ligation products by size or hybridization. Pr ...

Screening of recombinants It is best to do a test ligation without insert if the background is low it would not be necessary to screen colonies for insert.1) colony PCR1) Pick a colony from plate and ...

Stable Transfection (Electroporation)Outline:Electroporation can be used for both transient and stable transfection of mammalian cells. Cells are placed in suspension in an appropriate electroporation ...

基因组DNA Southern杂交1）基因组DNA Southern印迹的制备预备1．用适当的限制性内切酶消化基因组DNA样品（10μg）。2．进行琼脂糖凝胶电泳。一般用0.7～1.0％的琼脂糖凝胶分离基因组DNA，它在1～15kb的范围内有较好的分辨率，可选用TBE或TAE缓冲液。琼脂糖凝胶电泳需在1V/cm的电压下进行，如果要分离片段大小相似的DNA带，应用较大的凝胶（20×25cm）。常用的 ...

Restriction Map and Multiple Cloning Site of pEGFP-N2. (Unique restriction sites are in color or bold.) The Not I site follows the EGFP stop codon. The Nhe I site cannot be used for fusions since it c ...

Map of pLP-EGFP-C1 Vector. Unique restriction sites are shown in bold. ...

Restriction Map and Multiple Cloning Site of pbgal-Control. Unique restriction sites are in bold. ...

Restriction Map and Multiple Cloning Site of pbgal-Basic. Unique restriction sites are in bold. ...

Isolation of DNA from Acrylamide Gels1. 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 ...

Oligonucleotide PurificationSteve HahnLast modified 10/16/98Purpose: Purification of oligonucleotides (which have already been purified by reverse phase) can increase the efficiency of site directed m ...

DNA extraction from Mutation Detection Enhancement (MDE) Gel Stained with Silver NitrateSource: Contributed by Mohammad Reza Abbaszadegan et al.Abstract: Single strand conformational polymorphism (SSC ...

Electro-elution of nucleic acids from agarose and polyacrylamide gel slides.PrincipleThe nucleic acids contained within the gel slice are electrophoresed out of the gel funneled down into the v-shaped ...

Electro-elution of nucleic acids from agarose and polyacrylamide gel slides.PrincipleThe nucleic acids contained within the gel slice are electrophoresed out of the gel funneled down into the v-shaped ...

Recovering DNA from agarose gels Paul N. Hengen Ph.D. (July 14 1999) Introduction========Methods and reagents is a unique monthly column that highlights current discussions in the newsgroup bionet.mol ...

Materials: TE solution 10 mM Tris (pH to 7.5) 1 mM EDTA (pH to 8.0 to dissolve)Frozen agarose gel piece containing the desired DNA fragmentSupplies: Micropipetter and tips Microcentrifuge and tubes Sp ...