...during the scraping process. Add each scrape into a 50 mL conical tube on ice pooling the scrapings of two...spreader and cold LB to scrape bioassay plates and remove bacteria. Use one scraper for all plates. Use two...mL cold LB to each plate for each scrape and use spreader to scrape plates. Pro-Tip Pushing motion is ... each tube on ice while scraping. Repeat addition of 10 mL cold LB and scrape for each plate until agar...plasmids in a single sample. They are often used for screening, barcoding, or other high throughput multiplexed...activating pulse and follow all specifications described in the equipment manual. Immediately add 1 mL ...µl aliquots of cell/DNA mixture used (8 in the described protocol) using a new vented falcon tube for every...

...transforming into bacteria D. Screening for Inserts D.1 Recommended materials D.2 Screening for inserts E. Producing...10878/ . Description Vector Element U6 Human U6 promoter drives RNA Polymerase III transcription for generation...non-hairpin insert. pLKO.1 – scramble shRNA Negative control vector containing scrambled shRNA. psPAX2 Packaging...greater yield of DNA. Back to Top D. Screening for Inserts You may screen for plasmids that were successfully...generation of shRNA transcripts. cPPT Central polypurine tract, cPPT, improves transduction efficiency ...the pLKO.1 TRC-cloning vector. Addgene Plasmid Description pLKO.1 – TRC control Negative control vector ...R0101S NcoI NEB #R0193S Agarose Sigma #A9539 D.2 Screening for Inserts Day 1: 1. Inoculate 5 colonies from...

...#52962 ) and is described in Improved vectors and genome-wide libraries for CRISPR screening. Sanjana NE,...This protocol describes how to generate a monoclonal cell line from a polyclonal pool of stable cells...Protocols Viral Service Introduction This protocol describes how to generate a monoclonal cell line from a ...type will be able to grow under the conditions described in this protocol. Conditioned medium (details ... specific conditions used. Each 96-well plate described in this protocol could, in theory, give rise to...protocol was performed at Addgene on the A549 cells described in our sample data , each 96-well plate gave rise...monoclonal lines have been sufficiently expanded, screen the lines for transgene expression and/or other...

...collection of Addgene's video content, including how-to screen captures, lab procedure protocols, and career advice...standard laboratory procedures. How-To Videos Screencapture walk-throughs to help guide you through Addgene's...Protocols (Link opens in a new window) Video Link Description Related Page Making LB Agar Plates Create plates...Videos (Link opens in a new window) Video Link Description Related Page MTA Guide - Requests Your guide ...Videos (Link opens in a new window) Video Link Description Jessica Welch, PhD In this first installment ...

...antibody applications. Intro to the Lab Bench Name Description (Link opens in a new window) Link to Video Personal...materials on a balance Basic Molecular Biology Name Description (Link opens in a new window) Link to Video Making...without a commercial kit Plasmid Cloning Name Description (Link opens in a new window) Link to Video Restriction... targeting human and mouse genes. Virus Name Description Link to Video General Transfection Introduce ... your lentivirus preparation Antibodies Name Description Link to Video Transfection Introduce plasmid ...

...This protocol describes how to affinity purify recombinant antibodies from cell culture supernatant using...Western Blot Protocol Introduction This protocol describes how to affinity purify recombinant antibodies ... the reservoir to a LoBind tube. Pro-Tip Do not scrape the filter with the pipette tip. Determine the ...

...This protocol describes how to run an indirect ELISA against a purified antigen to demonstrate the expected...Protocol Antibody Guide Introduction This protocol describes how to run an indirect Enzyme-Linked ImmunoSorbent...antigen to validate an antibody. This protocol will describe how to create a standard curve using known concentrations...

... Biology Reference Introduction This protocol describes how to use droplet digital PCR (ddPCR) to titer...the 96-well plate sealer by gently touching the screen. Prepare the Serial Dilution Place a 48-well dilution...the plate sealer is not at temperature, touch the screen on the plate sealer to allow it to get to temperature...

...Guide Virus Blog Posts Introduction This protocol describes how to use droplet digital PCR (ddPCR) to titer...the 96-well plate sealer by gently touching the screen. Preparation of the Master Mix Place a ddPCR plate...the plate sealer is not at temperature, touch the screen on the plate sealer to allow it to get to temperature...

...overnight culture to 500 μL of 50% glycerol in a 2 mL screw top tube or cryovial and gently mix. Notes: Make...use a sterile loop, toothpick or pipette tip to scrape some of the frozen bacteria off of the top. Do ...

...bath on and set the appropriate temperature as described in your protocol. How to set the temperature will...water bath even if you are using disinfectants as described above in step 3. You will also need to maintain...

...This protocol describes the basic steps for fixing and staining cells in culture with a primary antibody...antibodies to detect antigens in cells. Here we describe the basic steps for fixing and labeling cells ...

...resolution (the ability to distinguish between two discrete objects). Figure 1 depicts an image of a compound... sure to take care of your microscope. Bumps, scratches, and dust can easily impair microscope performance...

...#52962 ) and is described in Improved vectors and genome-wide libraries for CRISPR screening. Sanjana NE,...

...This protocol describes the basic steps for lysing cells, determining total protein concentration in ...specific to the protein of interest. This protocol describes the basic steps for lysing cells, determining ...

... Bacterial Culture Introduction This protocol describes methodology for plating antibiotic over-agar for...

...Autoclave and allow to cool to room temperature. Now screw on the top of the bottle and store the LB at room...

...Protocols Viral Service Introduction This protocol describes a general method for transfecting mammalian cells...

...make your own. Transform bacteria with the DNA and screen for the correct plasmid product by restriction ...

...gene of interest in cell culture. This protocol describes how to transfect suspension HEK293 cells with ...