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...lots of FBS can promote or inhibit transfection. Test a variety of brands and lots of FBS to find one ...inactivated in the lab by heating to 56 °C for 30 minutes. 0.45 μm polyethersulfone (PES) filter system, ...and add 35 mL of 0.05% Trypsin/EDTA. Wait ~2-3 minutes for cells to detach. Gently tap the sides of the... and sonicate 5 x 1 sec pulses with at least 5 minutes on ice between each pulse, 50% amplitude. Return...

...mixes and samples on ice before use. Wipe down all pipettes and surfaces with 10% bleach. Safety Warnings ...PCR tubes, Axygen, PCR-02-FCP-C ddPCR 96-well PCR plates, Bio-Rad, 12001925 Pierceable foil heat seal, Bio-Rad...Cell Harvest Gently aspirate media from the 6-well plates. Be sure to use a new aspirating pipette for each...and Troubleshooting We recommend wiping down all pipettes and equipment with 10% bleach prior to use and...

...with standard agarose gel electrophoresis, which separates DNA by their length in base pairs. Following electrophoresis...Electrophoresis Protocol with the following amendments: Notes: Gel purification is most efficient with lower %...blade, slice the desired DNA fragment from the gel. Notes: To protect the UV box, it is a good idea to place...

...commonly used to culture bacteria in the lab. LB agar plates are frequently used to isolate individual (clonal...characterized by a cloudy haze in the media (Figure 1). Notes: Some protocols require bacteria to be in the log...on your plasmid. If the bacteria on your LB agar plates are not fresh, you should streak your bacteria ...

...Gibson Cloning? No need for specific restriction sites. Join almost any two fragments regardless of sequence...unique function for the reaction: T5 Exonuclease - creates single-strand DNA 3’ overhangs by chewing back ...other. Phusion High-Fidelity DNA Polymerase - incorporates nucleotides to “fill in” the gaps in the annealed...

...restriction sites. See (Link opens in a new window) NEB's table of methylation sensitive restriction sites . Sometimes...similar, but not identical, to their recognition sites. This is due to "Star Activity" and can happen ...

...lots of FBS can promote or inhibit transfection. Test a variety of brands and lots of FBS to find one ... plate in DMEM complete in 10 cm tissue culture plates. Incubate the cells at 37 °C, 5% CO 2 for ~20 h...the table below for a possible range of ratios to test: Ratio of DNA:PEI Amount of DNA (μg) Volume of 1...

...ultracentrifugation. The arrow indicates the 60–40% interface. The vertical black line indicates the location of the...fractions once you have a good idea when the virus elutes from the gradients. In a biosafety cabinet, carefully...

...of our protocols supports reproducibility and accelerates science. Here, we list the specific equipment...tubes, VWR 76289-498 Before Starting Wipe down all pipettes and equipment with 10% bleach prior to use. Warm...Spectrophotometer using the A280 IgG setting. Combine all eluates with measurable protein. Determine the protein ...

...of our protocols supports reproducibility and accelerates science. Here, we list the specific equipment...January 24, 2022 Workflow Timeline Day 1: Prepare lysates, run SDS-PAGE, transfer, block, incubate with primary...channel pipette Pipette controller Pipette tips and pipettes Spectrophotometer Heat block Mini gel tank chamber...

...Ready to learn about antibodies? Our latest animation is here to help! Join Abi as they explain what ...

...on the surfaces of the water bath as the water evaporates from the tub. Disinfectant may be added to prevent...heat up quicker. Pro-Tip Set up the water bath 30 minutes to 1 hour before you need to use it to allow the...

...determine your antibody's purity? Check out our latest protocol video, "Coomassie Purity Stain of Recombinant...

...where to start or what your options are? Addgene's latest animation is here to help! "Antibodies 101: Antibodies...

...We are very excited to share the latest edition of our Viral Vectors 101 eBook!  This practical guide...

...regularly from now until May, so follow us to get updates whenever a new video is posted! If TikTok isn't...

...some cheer by snapping a photo of you and your labmates celebrating the holidays and tweeting it with ...

...UPDATE: The contest is now closed. Stay tuned to see the finalists and vote for your favorite! Our mascot...

... 0.5-1μL T4 DNA Ligase H 2 O to a total of 10μL Notes: If the DNA concentrations are low such that you...troubleshooting failed ligations. The following table indicates the various controls: Control Ligase Interpretation...

...of our protocols supports reproducibility and accelerates science. Here, we list the specific equipment... Equipment Pipette controller Pipette tips and pipettes Rocking platform Tweezers Fluorescent microscope...