...enzyme digest (subcloning), including design and experimental procedures.... Protocols Subcloning Plasmid Cloning...Cloning by Restriction Enzyme Digest (aka Subcloning) You may also like... Restriction Digest of Plasmid...out our Empty Backbone Reference . Background Subcloning by restriction digest is a commonly used lab ...YGOI is in a bacterial expression vector. Using subcloning, you can easily move YGOI into a mammalian expression...find two different restriction enzymes for your subcloning. It is also possible to use a single enzyme, ...

... of a substance in that liquid. In order to accurately measure the concentration of a substance based ...to determine the concentration of a particular substance in that liquid. Molecules absorb different wavelengths...need to know the wavelength of light that your substance maximally absorbs. In the case of nucleic acid...

...powder 96-well microtiter plate Chemiluminescence substrate Plastic wrap Primary antibody Secondary antibody...absorbance of the duplicate samples on the plate. Subtract the average absorbance of the blank from all of...shaking platform. Prepare the chemiluminescence substrate by mixing 1:1 reagent A to reagent B. Gently incubate...

...,5'-tetramethyl-[1,1'-biphenyl]-4,4'-diamine) substrate kit, Pierce 34021 Tween-20, VWR 0777-1L Foil Before...Solution and 5 mL Peroxide Solution from the TMB substrate kit. Cap the tube and invert several times to ...secondary antibody for 2 hours, reacted with TMB substrate and the reaction stopped after 15 minutes. Absorbance...

...function. We specifically use this protocol with a subclone of HEK293T cells optimized for viral production...Workflow Timeline Day 0: Seed HEK293T cells (or a subclone of HEK293T optimized for viral production) Day...

... lower the pipette tip into the liquid. Do not submerge the pipette itself into the liquid and avoid touching...contaminating the contents. Once the end of the tip is submerged, slowly release the plunger, allowing it to go...

... a water bath at 60 ℃ with sufficient water to submerge ~75% of the bottle containing your molten gel ...removing anything from the autoclave. Partially submerge your molten gel-mix in the 60 ℃ water bath. Pro-Tip...

... thought on using selective medium throughout subsequent experiments. When selective medium is used, it...because the buildup of waste products could be suboptimal for cell growth. This conditioned medium will...

...can be converted into lentiviral particles for subsequent infection of a target cell line. Once introduced...toxic to some cell lines. In these cell lines, substitute protamine sulfate for polybrene. c. Add lentiviral...

...stable for years, as long as it is kept at -80°C. Subsequent freeze and thaw cycles reduce shelf life. To ...

...with an empty tube. Alternatively, you can just subtract the weight of the empty tube from the weight of...

...weighing and prevent cross-contamination with other substances. They also help you transfer the material to ...

...buffer) to ensure optimal salt concentrations in subsequent reactions. Step 3: Create Vector Overhangs Treat...

...vary between cell lines. It is not recommended to subject lentiviral supernatants to multiple freeze-thaw...

... DNA 5’ end. Complementary DNA fragments can subsequently anneal to each other. Phusion High-Fidelity ...

...not recommended that lentiviral supernatants be subjected to multiple freeze-thaw cycles. This protocol ...

...not recommended that lentiviral supernatants be subjected to multiple freeze-thaw cycles. Procedure Before...

...the standard curve is y = 14725x + 3748.4, then substitute 18179.442 for y and solve for x. 18179.442 = ...

...centrifuge with the mark facing outward in all subsequent centrifugation step Pro-Tip Resin will appear...

...incremental, ordered cell lysis. E. coli cells are subject to lysis by freeze-thaw if not suspended in cryoprotectant...