We narrowed to 50 results for: URE

...glycerol stocks. Procedure Follow the steps for Inoculating an Overnight Liquid Culture . After you have... appropriate temperature. Growth conditions, including copy number and growth temperature, can be found...you retrieve your liquid bacterial culture, take 500 μL of culture to make your glycerol stock before ...times). Make sure that you see one uniform solution, and there are no layers present. Be sure to label both...Transformation Recovering Plasmid DNA from Bacterial Culture Background Information Bacterial glycerol stocks... bacterial growth, add 500 μL of the overnight culture to 500 μL of 50% glycerol in a 2 mL screw top tube...next day you will be able to start an overnight culture for plasmid DNA prep the following day. Tips and...

...cm tissue culture plates. Incubate the cells at 37 °C, 5% CO 2 for ~20 h. Prepare a mixture of the three... 113.4 Gently add the diluted PEI mixture to the diluted DNA mixture. Add the diluted PEI dropwise while...polyethylenimine (PEI) transfection protocol. This procedure can be modified for alternative packaging cell...Microcentrifuge tubes, Neptune 3745.X 10 cm tissue culture dish, Corning 430167 15 mL conical tubes, VWR 21008...solution to mix for 10 min and then recheck the pH to ensure that it has not drifted. Filter the solution through...that are below passage 15 for viral production. Procedure Seed 293T packaging cells at 3.8×10 6 cells per...negative E. coli strain such as NEB stable. Make a mixture of a total of 500 μL PEI-OptiPro SFM with enough...

... counter and measure the live cell density and viability of the culture. Pro-Tip Culture should be between...20 °C. Procedure Section 1: Seeding cells The day prior to transfecting, seed a 108 mL culture of HEK293...transient expression of a gene of interest in cell culture. This protocol describes how to transfect suspension... cells are considered biosafety level 2. Please ensure that you are in compliance with your institution...the DNA and working stock of PEI-MAX to room temperature before use, and warm the BalanCD HEK293 transfection...water to a final volume of 1 L and recheck pH to ensure that it has not drifted. Filter the solution through...biosafety cabinet and incubate for 1 h at room temperature. After the BCD TFX has warmed in the bead bath...

...samples at specific temperatures. This protocol will cover the general procedure and features to keep in mind...densities is essential to many different experimental procedures, such as minipreps and RNA extractions. Plus,... centrifuge may have specific instructions so be sure to check in with someone familiar with your lab’...the time and speed needed for your experiment. Procedure An example of a balanced tabletop centrifuge, ...a tabletop microcentrifuge like the one in the picture above. Wear PPE appropriate for the lab space in.... Before using the centrifuge for your samples, ensure that the centrifuge is clean and that everything...center (see image to right). Even distribution ensures that the centrifuge is balanced. Using the centrifuge...

...Biosafety Levels One and Two (BSL-1 and BSL-2) Safety measures for laboratories operating at BSL-1 and BSL-2 ...the building blocks for many more complicated procedures. Name Description (Link opens in a new window...to Video Making LB Agar Plates Create plates to culture bacteria in the lab Watch the Video! Over-Agar ...Antibiotic Plating Quickly add antibiotic to a pre-poured plate Watch the Video! Streaking Bacteria Isolate...Watch the Video! Inoculating a Liquid Bacterial Culture Prepare and grow bacteria in liquid medium Watch...extract, and precipitate DNA DNA Quantification Measure DNA concentration with a spectrophotometer Restriction...Video! Sequence Analysis Verify important plasmid features using sequence analysis RNA Extraction Without...

...and click measure, be sure to record the concentration and purity. Note: Purity is measured under the ...Inoculating a Liquid Bacterial Culture Recovering Plasmid DNA from Bacterial Culture Restriction Digest of Plasmid...substance in that liquid. In order to accurately measure the concentration of a substance based on its absorbance... of whether you have a NanoDrop, follow the manufacturer's instructions for the spectrophotometer specific...Spectrophotometer Tips Before measuring any samples, be sure to ‘blank’ the spectrophotometer using the solution...resuspended in, but with no DNA added. 'Blanking' measures the background inherent to the machine and your...your solvent. If using a NanoDrop to measure your samples, place 1-2µL of mini-prepped DNA onto the pedestal...

...list above. Procedure Option #1 - Solution D Protocol Before starting this protocol, make sure to prepare...TRIzol® User Guide from ThermoFisher Scientific . Figure 1: A diagram of the different steps in RNA extraction...step, Option B) Glycogen (Optional) Caution Make sure to read the SDS (Safety Data Sheet) for safety warnings...use 1 mL of Solution D per 100 mg of cells. For cultured cells: use 1 mL of Solution D per 1 X 10 7 cells...cells. Allow sample(s) to sit at room temperature for 5 minutes to allow for dissociation of the nucleoprotein...centrifuge, bring this equipment back to room temperature, as prolonged storage in the cold room may damage...top, aqueous phase to a new RNAse-free tube. The mixture separates into a bottom organic layer, an interphase...

...DNA from Bacterial Culture Introduction If you have a glycerol stock or stab culture of bacteria and need...inoculate a bacterial culture for DNA purification will minimize the chance of having a mixture of plasmids in...touch the bacteria growing within the punctured area of the stab culture or the top of the glycerol stock.... (with appropriate antibiotic) Bacterial stab Procedure Obtain an LB agar plate with appropriate antibiotic...sterilize it by passing it through a flame, just be sure to allow enough time for the loop to cool before...diagonal lines in another section of the plate. Make sure that the first line (and only the first) in each...plasmids, which sometimes recombine at 37 °C. Be sure to check this before incubating your plate. In the...

...reagents for a stock mixture is an essential laboratory technique, as imprecise measurements can affect the ...other solutions. A key part of this task is making sure you’re weighing all reagents precisely to create....) Reagents Material that will be weighed out Procedure The balance in the photo above has a capacity ...need to weigh it out in the fume hood to prevent exposure. Read the reagent’s material safety data sheet...by looking for a weight range on the scale. Make sure that the weight of the material you’re weighing ...weight of the weighing boat or paper into your measurement. Hit the tare button (or zero button) to set ... material directly to the tube to weigh it. Make sure that your scoopula or spatula is clean, and sterile...

...New England Biolabs B7022S Procedure Pouring a Standard 1% Agarose Gel: Measure 1 g of agarose. Pro-Tip ...pipette tip. Place newly poured gel at 4 °C for 10-15 mins OR let sit at room temperature for 20-30 mins, until...Introduction Gel electrophoresis is the standard lab procedure for separating DNA by size (e.g., length in base...very little difference between the two. Note: Make sure to use the same buffer as the one in the gel box...earlier so that it is already cold when the gel is poured into it. Loading Samples and Running an Agarose...loading the sample in the well, maintain positive pressure on the sample to prevent bubbles or buffer from...DNA ladder in the first lane as a guide (the manufacturer's instruction will tell you the size of each ...