...(CRISPRi) or an activation domain (CRISPRa) to regulate gene expression. Some CRISPR libraries contain...

...tendency to runout right when you need them and to accumulate a bunch of unused buffers because you’ve run ...

...helpful information about the subject, link to related guides and protocols, and provide easy to sort ...

... deliver a large amount of DNA (~8 kb) with a relatively low immune response, it should come as no surprise...

... yourself a Scientist. You work in a lab. You formulate hypotheses and conduct experiments. You transfer...

...that’s only good once. Even when things return to relative normalcy, I’ll be incorporating that mobile monitor...

...cells with plasmids encoding aptamer-tagged RNA and later introducing the small molecule into the cell. The...

...courses on technologies and laboratory techniques related to antibody discovery. You can find cell surface...

...most commonly used in the laboratory for genome manipulation. When transposons are used in the lab, the transposase...

...’re working with and better understand how to manipulate it. Happy CRISPRing!   Molecular graphics and...

...overnight. Plate 2.5 mL of the transformed cells on each of the eight bioassay plates (two plates per tube...Antibiotic 245 mm bioassay plates (Molecular Devices, X602) Pro-Tip Pour these plates at least one day in advance... Agar + Antibiotic plates at 37 °C. Prewarm 8X LB Agar + Antibiotic Bioassay plates. Prechill Micropulser...100 dilution). Plate 100 µL of each dilution onto a prewarmed Petri dish. Incubate plates at 30 ℃ overnight... edge that can scrape plates more abrasively at a certain angle. Incubate plates upside down at 30 ℃ overnight... are not present on the most dilute plate, count the second plate (ie. the 1:10,000 dilution). After 12...LB to scrape bioassay plates and remove bacteria. Use one scraper for all plates. Use two 10 mL pipettes...

... forget to factor this into your calculations later (see calculation example below). Seed 300,000 cells...well plate sealer by gently touching the screen. Preparation of the Master Mix Place a ddPCR plate onto... Foil Heat Seal on the PCR plate with the red line facing up. If the plate sealer is not at temperature...place the PCR plate with the foil onto the metal support block. Place the block in the plate sealer and ...transfer the plate to the Droplet Reader. Open the QuantaSoft software to set up a new plate layout. Designate... a CSV file which will be used to calculate the titer. Calculations In the experimental setup above, the...dilution into 6-well plate 2.5 25 5 50 10 100 20 200 40 400 80 800 160 1600 To calculate the titers, first...

...02-FCP-C ddPCR 96-well PCR plates, Bio-Rad, 12001925 48-well dilution plate, Bio-Rad, MLL4801 Pierceable... Foil Heat Seal on the PCR plate with the red line facing up. If the plate sealer is not at temperature...place the PCR plate with the foil onto the metal support block. Place the block in the plate sealer and ...transfer the plate to the Droplet Reader. Open the QuantaSoft software to set up a new plate layout. Designate...compliance with your institution’s biosafety regulations. Equipment Class II, Type A2 Biological Safety...Bio-Rad, DX200 Thermal Cycler, Bio-Rad, T100 PCR Plate Sealer, Bio-Rad, PX1 1–10 µL single channel pipette...reagents and the NTC with virus. Pre-warm the 96-well plate sealer by gently touching the screen. Prepare the...

...standards and samples onto the qPCR plate Set up and load the 96-well plate: Load 5 μL of each standard in ... 58 °C 30 sec / read plate/ repeat 39x from step 3 / melt curve Example of plate set-up: 1 2 3 4 5 6 7...these institutions. This protocol is for a 96-well plate with 20 μL reaction volume. Last Update: February...Estimate of time required: ~3 h Workflow Timeline Plate set-up: 2 h qPCR run: 1.5 h Data analysis: 30 min...min Equipment qPCR instrument Heating plate Pipettors 1–10 µL single channel pipette 20–200 µL single ...Nuclease-free water Microcentrifuge tubes 96-well optical plate Pipette tips General Considerations Always run standards... the samples being tested. Always include a No Template Control (NTC), i.e master mix + water Whenever...

... We recommend that you plate 50 μL on one plate and the rest on a second plate. This gives the best chance...they will be able to grow once plated on the antibiotic containing agar plate. This step is not critical ...antibiotic resistances. Plate some or all of the transformation onto a 10 cm LB agar plate containing the appropriate...isn't too much liquid media on the agar plates. If the agar plate doesn't dry adequately before the cells... What went wrong? Check that you are plating on an LB Agar plate containing the correct antibiotic. The...also make their own competent cells. This is a relatively simple procedure and is useful for performing...Watch the protocol video below to learn how to isolate single bacterial colonies. Equipment Shaking incubator...

...cells, following manufacturer’s protocol. Plate on LB agar plates containing 100 μg/mL ampicillin or carbenicillin... transfected, plate 7×105 HEK-293T cells in 5 mL of media in a 6 cm tissue culture plate. Incubate cells...initiating your experiment. Day 1: a. Plate target cells in ten 6 cm plates and grow at 37°C, 5% CO 2 overnight...that you maintain one uninfected plate of cells in parallel. This plate will serve as a positive control... A.1 The RNAi Consortium A.2 Map of pLKO.1 A.3 Related plasmids B. Designing shRNA Oligos for pLKO.1 B...terminal repeat. RRE Rev response element. A.3 Related Products The following plasmids available from ... have at least 3 nucleotide mismatches to all unrelated genes. TIP: Addgene recommends that you select...

...window) Video Link Description Related Page Making LB Agar Plates Create plates to culture bacteria in the...the lab Pouring LB Agar Plates Protocol Streaking Bacteria on Plates Isolate single bacterial colonies ...Antibiotic Plating Create plates to culture bacteria in the lab Over-Agar Antibiotic Plating Protocol AAV...colonies on an agar plate Streaking Bacteria on Plates Protocol Diagnostic Restriction Digest Verify a plasmid...efficiency Bacterial Transformation Protocol Inoculating a Liquid Bacterial Culture How to prepare and...and grow bacteria in a liquid medium Inoculating a Liquid Bacterial Culture Protocol Getting Started with... an Agarose Gel Gel purification allows you to isolate and purify DNA fragments based on size. Following...

...If you are using the same template DNA for all your reactions, the Template DNA can be added to the master...particularly for GC-rich templates. What does each ingredient specifically do? Template DNA: Contains the portion...amplifying for analysis and manipulation. It is essential that the template DNA and primer are complementary...Taq Polymerase. From a single copy of DNA (the template), a researcher can create thousands of identical...polymerases from other species. In addition to the template DNA and the Taq polymerase, PCR requires free ...micrograms of target DNA from just a few nanograms of template DNA through several cycles of denaturation, annealing... initiation step heats the double stranded DNA template strand to the point where the strands start denaturing...

... from different cells in the population. Once the polyclonal populations are growing well and have been...Consider expanding populations transduced with a variety of dilutions and pick the population that has the ...days. Single cells were then plated in individual wells of a 96-well plate and left undisturbed for 13 ...a more homogenous (but still polyclonal) cell population. Depending on the transducibility of the cell...selection may be a vital step for obtaining a population of cells that have taken up the lentiviral transgene... well (each well gets one dilution, so a 6-well plate will hold 5 dilutions plus one 'no virus' control...be 10 µg/mL. Pro-Tip Transducing too many cells relative to the number of virus particles reduces the transduction...

...rocking platform. Permeabilize cells for 10 min at room temperature ( RT ) on a rocking platform in 500...rocking platform. Section 3: Labeling with antibody Block for 20 min at RT on a rocking platform in 500...Pipette controller Pipette tips and pipettes Rocking platform Tweezers Fluorescent microscope 0.5–10 µL single...Poly-D-lysine coated coverslips HeLa cells 24-well plate 4% Paraformaldehyde 5 mg/mL 4′,6-diamidino-2-phenylindole... in each well of a 24-well cell culture treated plate. Seed 5 x 10 3 HeLa cells per well. Allow the HeLa...cells Gently aspirate the media from the 24-well plate. Wash each well with 500 µL of PBS, remove the wash.... Wash 3x for 5 min in 500 µL PBS on a rocking platform. Dilute the fluorescently-labeled secondary antibody...