...Creating Bacterial Glycerol Stocks for Long-term Storage of Plasmids Creating Bacterial Glycerol Stocks for Long-term...time, you will need to establish glycerol stocks. The addition of glycerol stabilizes the frozen bacteria...of 50% glycerol in a 2 mL screw top tube or cryovial and gently mix. Notes: Make the 50% glycerol solution...concentration of long-term glycerol storage is unknown. Most labs store bacteria in 15-25% glycerol. You can prepare...your glycerol stock before you begin your plasmid mini-prep. Try not to freeze/thaw your glycerol stock...Protocol for creating a glycerol stock, including additional tips for long-term storage. Protocols...Bacterial Culture Background Information Bacterial glycerol stocks are important for long-term storage of ...

...how to generate a monoclonal cell line from a polyclonal pool of stable cells....how to generate a monoclonal cell line from a polyclonal pool of stable cells. Transducing cells with ...with lentivirus results in a heterogenous polyclonal population that varies in the number of integration ... as the lower expressing clones take over the polyclonal cell pool. Generating a monoclonal cell line ... Hemocytometer or other cell counter Reagents Polyclonal stable cell pool DMEM high glucose, Corning 10...FBS and 5 mL of 100X glutaGRO. Store at 4 °C. Polyclonal stable cell pool: see our protocol for generating...cell lines with lentivirus . Pro-Tip Because the polyclonal cell pool is always undergoing selection, it’...

...cells, resulting in a more homogenous (but still polyclonal) cell population. Depending on the transducibility...regularly to confirm transgene expression. As polyclonal populations of resistant cells start proliferating...etc. Pro-Tip This selection method results in a polyclonal cell population, meaning that the transgene has...from different cells in the population. Once the polyclonal populations are growing well and have been sufficiently...expression. Over time, transgene expression in a polyclonal population may drop. This is because cells that...consider generating monoclonal lines from the early polyclonal populations. Sample Data Figure 1: Generation...

...from Bacterial Culture Introduction If you have a glycerol stock or stab culture of bacteria and need to ...burner. Obtain the approrpriate bacterial stab or glycerol stock . Using a sterile loop, pipette tip or toothpick...punctured area of the stab culture or the top of the glycerol stock. Pro-Tip If you use a wire loop you can ...

...Lithium Chloride (for precipitation step, Option B) Glycogen (Optional) Caution Make sure to read the SDS (...anticipate your RNA yield to be small, RNase-free Glycogen may be used as a carrier to facilitate RNA precipitation...

...Bacteria on an LB Agar Plate Creating Bacterial Glycerol Stocks for Long-term Storage of Plasmids Recovering... the bacteria, you can proceed with Creating a Glycerol Stock . You can now isolate your plasmid DNA from...

...liquid medium Watch the Video! Creating Bacterial Glycerol Stocks Store bacterial strains or plasmids for...Dilution Generate monoclonal cell lines from a polyclonal pool of stable cells AAV Production in HEK293...

...Benzonase/DNAse I (Millipore 71205-3) 40% Polyethylene Glycol 8000 (PEG) + 0.5 M NaCl Cell lysis buffer (50 mM... and thaw a new working stock. 40% POLYETHYLENE GLYCOL (PEG) 8000 solution: Dissolve 400 g of PEG 8000...

...happen for a variety of reasons, including high glycerol concentration. Learn more at (Link opens in a ...

... migrated; 2) it contains a high percentage of glycerol that increases the density of your DNA sample ...

...freeze-thaw if not suspended in cryoprotectant like Glycerol or DMSO solutions. Quantify the individual Maxipreps...

...complex in the transduced cells. hPGK Human phosphoglycerate kinase promoter drives expression of puromycin...