DNA基础知识

Preparation of Electrocompetent Cells and Electroporation of Plasmid DNA Inoculate 5-ml L-broth with a single colony of E. coli. Incubate 5 hours to overnight at 37°C on a roller or with moderate shak ...

FROZEN COMPETENT E. COLI CELLS(Inoue et al. 1990 Gene 96:23) Inoculate a 5ml overnight of E.coli in LB+20 mM MgSO4. Next morning inoculate 250 ml LB+20 mM Mg++ in a 2L flask with about 2ml overnight c ...

Transformation "Ultra-Competent" E. coli (Inoue Method)Inoue H. H. Nojima and H. Okayama. 1990. High efficiency transformation of Escherichia coli with plasmids. Gene 96:23-28. Citation Abstract Proce ...

Extraction from Cheek Cells DNA

Just follow these 3 easy steps:DetergenteNzymes (meat tenderizer)AlcoholIt's that simple? Tell me more!First you need to find something that contains DNA. Since DNA is the blueprint for life everythin ...

Rubidium Chloride method for Transformation Competent E. coliProcedure1. Inoculate 1 ml from overnight culture into 100 ml Psi broth (scale up or down as needed). Incubate at 37 C with aeration to A55 ...

Recipe for yeast tryptone medium Keywords: Zappe; YT 2x; Yeast tryptone; E. coli Yeast tryptone medium Contributor: Zappe H Related: E. coli growth Yeast tryptone broth/agar 2xYT Component stock con ...

Recipe for yeast tryptone medium Keywords: Zappe; YT 2x; Yeast tryptone; E. coli Yeast tryptone medium Contributor: Zappe H Related: E. coli growth Yeast tryptone broth/agar 2xYT Component stock con ...

Isolating DNA Fragments Materials:• 0.8 % agarose gel in 1x TAE• Digested DNA• Glass Milk• NaI solution• New WashProcedure:1) Run digested DNA out on agarose gel slowly (70 V on BioRad gel)2) Use long ...

Gene-Kleen protocolPurification of DNA from agarose gels is an essential method involved in the sub-cloning of DNA fragments. The following method describes a variation of the method of Vogelstein and ...

CaCl2 Competent Cells Description: Competent cells are able to take up DNA.... Competent cells CaCl2 method E. coli long protocol Keywords: competent cells; E. coli; calcium chloride; Zappe CaCl2 com ...

CaCl2 Competent Cells Description: Competent cells are able to take up DNA.... Competent cells CaCl2 method E. coli long protocol Keywords: competent cells; E. coli; calcium chloride; Zappe CaCl2 com ...

Phenol-based Method for the Isolation of DNA Fragments from Low-Melting Temperature AgaroseReference: Favre D. 1992. Biotechniques vol. 13Cut out slice containing DNA smallest size possible. Estimate ...

ISOLATION OF DNA FROM AGAROSE GELS WITH DEAE PAPER MATERIALS:The DEAE paper can be used as supplied by Schleicher and Schuell however the binding capacity can be increased by washing the paper in 10m ...

ISOLATION OF DNA FROM AGAROSE GELS WITH DEAE PAPER MATERIALS:The DEAE paper can be used as supplied by Schleicher and Schuell however the binding capacity can be increased by washing the paper in 10m ...

Isolation of DNA from Agarose Gels (Paper Slurry Method) This procedure isolates DNA from agarose gels by filtration through a filter-paper column. The column is made in a 500 µL tube from a slurry of ...

An inexpensive alternative to glassmilk for DNA purificationPreparation of Silica1. Suspend 5 g of silica (Sigma S-5631) in 50 ml of PBS. 2. Allow the silica to settle for 2h. 3. Discard the supernat ...

Electroelution of agarose fragmentsElectroelution buffer1 M Tris pH 7.5 12.0 mls0.5 M EDTA 0.24 mls1 M NaCl 3.0 mlsqs to 600 mls dH2OAcetate cushion 3 M NaAcetate pH 4.8 480 ul0.1 % Bromphenol Blue 40 ...

Electroelution of agarose fragmentsElectroelution buffer1 M Tris pH 7.5 12.0 mls0.5 M EDTA 0.24 mls1 M NaCl 3.0 mlsqs to 600 mls dH2OAcetate cushion 3 M NaAcetate pH 4.8 480 ul0.1 % Bromphenol Blue 40 ...

DNA PrecipitationPhenol (removes protein)add equal volume of Phenol (= tris-saturated Phenol-Chloroform-Isoamyethanol) vortex spin 2 minutes at 12000 rpm 4°C transfer supernatant to a fresh tube (avoi ...