P3000™ reagent 0.5 μL 3 Add tube 2 solution to tube 1 and mix well 4 Incubate mixture from step 3 at room temperature for 10–15 min 5 Add 50 μL of complex from step 4 to cells; gently swirl plate to ensure homogeneous distribution of complex to the entire well ...
Tip: HepG2 cells tend to grow in clusters. It is critical to ensure dissociation has occurred to yield a single-cell suspension when passaging and plating the cells. Having a single-cell suspension is also very important to ensure efficient transfection....
3)Allow the solution in the HiPure Filter Midi Column to drain by gravity flow. 4)Prepare the cell lysate (see below) while the HiPure Filter Midi Column is equilibrating. 2.Preparing Cell Lysate 1)Harvest bacterial culture cells by centrifugation. For high copy number plasmids, harvest 15...
The plasmid DNA is washed/rinsed on the DNA-binding filter 8 using, e.g., an ethanol-containing wash solution such as 75% ethanol with 5 mM Tris-HCl, pH 7.4. The plasmid DNA may be eluted from the DNA binding matrix 8 in an appropriate aqueous buffer (e.g., TE or water) for im...
solution of a monovalent ionizable salt, and cationic liposomes comprising a cationic vesicle-forming vesicle lipid, dimethyldioctadecylammonium bromide (DDAB), N- [1- (2-3-ditetradecyloxy) propyl] -N, N-dimethyl-N-hydroxyethylammonium (DMRIE) bromide, N- [1- (2, 3-dioleyloxy) propyl-N,...
After incuba- tion at 37ЊC for 10 min, 50 l of cofactor solution (10 mM MgCl2, 5 mM CaCl2) was added and mixtures incubated at room temperature for further 10 min. Then, 2.5 × 10¹2 Kunitz units of DNase I (D-4263, Sigma) were added to each sample. After incubation at...
Vectashield mounting solution (Vector Laboratories, H100) was used to mount the samples, and fluorescence images were analyzed in detail using a confocal microscope (LSM700, Carl Zeiss, Germany). The signal intensities were quantified using ImageJ software (National Institutes of Health). 2.5. ...
3. The method of claim 1, wherein the pressure inside the muscle is increased by injecting PBS after plasmid DNA administration. 4. The method of claim 1, wherein the naked plasmid DNA is diluted in saline, PBS, sucrose solution, or glucose solution. 5. The method of claim 1, where...
After 1 week from the second inoculation, the mice are intraperitoneally immunized with 0.2 ml of a solution similar to the first one. Finally, after two weeks from the third inoculation a booster immunization is effected. The thus induced antibodies are collected in the ascitic fluid formed ...
PURPOSE: To increase the conversion into a 3-hydroxynitrile compound by making a culture solution of a specified transformed microorganism or its cell body (treated material) act on a 1,2-epoxy compound in the presence of an alkali cyanide.YAMADA HIDEAKI...