DNA实验技术

OUTLINE OF PROCEEDINGS USED IN TISSUE CULTURE TRANSFECTIONS1. Remove the required cells from liquid nitrogen and thaw at 37°C.2. Add the thawed cells to 10mls C2GM that has been equilibrated in the 3 ...

OUTLINE OF PROCEEDINGS USED IN TISSUE CULTURE TRANSFECTIONS1. Remove the required cells from liquid nitrogen and thaw at 37°C.2. Add the thawed cells to 10mls C2GM that has been equilibrated in the 3 ...

Electroporation of P. aeruginosa Preparation of Electrocompetent Cells Inoculate a 5-ml of L-broth with cells of the P. aeruginosa strain to be electrotransformed. Grow culture overnight at 37°C on a ...

Electroporation of P. aeruginosa Preparation of Electrocompetent Cells Inoculate a 5-ml of L-broth with cells of the P. aeruginosa strain to be electrotransformed. Grow culture overnight at 37°C on a ...

Materials:SOB medium E. coli host strain such as DH5αWBtRNA5 M ammonium acetate100% ethanol70% ethanol0.5X TESOC mediumtransformation platesI. Preparation of E. coli cells for electroporation.1. Use a ...

How to Make Competent Cells Step 1: Take an 100 mL aliquot of frozen cells from the -80oC and innoculate about 500 ml to 1 L sterile LB broth. DO NOT ADD antibiotic since these cells do not have an pl ...

Deproteination using phenol/chloroform (Maniatis et al. 1982)'Phenol' as used is Analar grade. Phenol should be melted at 65°C8-hydroxyquinoline added to a final concentration of 0.1% and equilibrated ...

Dialysis (using dialysis tubing)Molecularpourous membrane tubing is used for desalting protein and DNA isolation / purification. It comes in several diameters and pore sizes (for molecular weight conv ...

NH4Ac and EtOH precipitation of DNAAdd NH4Ac (10 M stock or solid) to the sample for a final concentration of 2.5 M mix (spin at 4 C transfer the supernatant to a new tube; optional spin for extra pur ...

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 ...