...and click measure, be sure to record the concentration and purity. Note: Purity is measured under the ...specific wavelength that they maximally absorb. By measuring the absorbance of a liquid you can accurately ...substance in that liquid. In order to accurately measure the concentration of a substance based on its absorbance...specific to your lab. Spectrophotometer Tips Before measuring any samples, be sure to ‘blank’ the spectrophotometer...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...

...Biosafety Levels One and Two (BSL-1 and BSL-2) Safety measures for laboratories operating at BSL-1 and BSL-2 ...extract, and precipitate DNA DNA Quantification Measure DNA concentration with a spectrophotometer Restriction...Lentivirus Quantify virus using fluorescence measurements Colony Formation Titering Assay for Lentivirus...by qPCR Using SYBR Green Technology Use qPCR to measure the physical titer of your adeno-associated virus...the Video! AAV Titration by ddPCR Use ddPCR to measure the physical titer of your adeno-associated virus...preparation Lentivirus Titration by ddPCR Use ddPCR to measure the physical titer of your lentivirus preparation...

... However, it is also more expensive. Procedure Measure 37g of pre-mixed LB-agar powder per L of molten...agar you’d like to make. The precise mass you measure out will be based on the number of plates you’d...case we spill anything or have small errors in measurement. You should also always make a bit more gel-ager...LB-agar powder. Transfer the LB-agar powder you’ve measured out into an appropriately sized bottle for autoclaving... solution of 100,000 ug/mL (100 mg/mL). Simply measure out 100 mg of ampicillin powder, add it to 1 mL...at a time next to the flame and begin pouring. Measure your desired amount of agar with a pipete for the...

...rocking platform in 500 µL permeabilization buffer. Remove the permeabilization buffer and dispose of it...antibodies raised in a mouse. Reagent Preparation Permeabilization buffer: Dilute 20 µL of Triton X-100 in 10...density before labeling. Section 2: Fixing and permeabilizing cells Gently aspirate the media from the 24...for 5 min in 500 µL PBS on a rocking platform. Permeabilize cells for 10 min at room temperature ( RT ) ...

...pipette for an accurate measurement in the next step. The second stop is meant to expel excess liquid ...Pipette tips Waste container Containers to hold measured liquid (ex: microfuge tube, bottle, etc.) Labels...tip is so that the same pipette can be used for measuring different samples without cross contamination ...tip into the container you are using to hold the measured liquid (ex: another microcentrifuge tube, a bottle...

...gel extraction kit. Elute in 30 μL of ddH 2 O. Measure the DNA concentration. C.4 Ligating and Transforming...TRC-cloning vector from step C.3 (If you were unable to measure the DNA concentration, use 1 μL) 2 μL 10x NEB T4...distribution of any literature by Addgene is not meant to carry with it, and does not grant any license...The distribution of materials by Addgene is not meant to carry with it, and does not grant any license...

...essential laboratory technique, as imprecise measurements can affect the outcome of your experiment. In...weight of the weighing boat or paper into your measurement. Hit the tare button (or zero button) to set ...

...bacteria but also because bacteria are used as the means for both storing and replicating plasmids. Because...applied to the cell/DNA mixture to induce membrane permeability. To do this you will need to have access to ...

...each other. When the sticky ends are compatible, meaning that the overhanging base pairs on the vector and...average size genes and vectors, this ratio is really meant to refer to the molarity of DNA ends available for...

... in healthy humans. Many of the precautions are meant to protect individuals from potential accidents ...

...provider. My sequence has “N”s in it- what does that mean? “N” is the letter used to denote that the position...

...your specific protocol and conduct an OD600 to measure the density of your culture if needed. A good negative...

...In our case, dGTP will be used in the reaction, meaning that T4 Pol will remove bases from the 3' end of...

...while working. AAV Titration by qPCR Use qPCR to measure the physical titer of your adeno-associated virus...

...different types of hazards and to provide protective measures for each level, as each level has different safety...

...ratios provided high transfection efficiencies as measured by the highest proportion of GFP positive cells...

...counting chamber on an automated cell counter and measure the live cell density and viability of the culture...

...) Procedure Pouring a Standard 1% Agarose Gel: Measure 1 g of agarose. Pro-Tip Agarose gels are commonly...

...150 ng/μL to several μg/μL. The protocol below is meant to describe the general procedure for purifying ...

...method results in a polyclonal cell population, meaning that the transgene has integrated in different ...