... Novel Next-Generation Sequencing and Analysis Platform to Assess the Identity of Recombinant Adeno-Associated...

...technique should I use for my experiment? When I am contemplating which microscope to use for a new sample I always...

.... “A Unified Multi-Kingdom Golden Gate Cloning Platform.” Scientific Reports, vol. 9, no. 1, Springer ...

... expand the self-labeling capabilities of this platform. The Janelia Fluor ligands provide brighter and...

... provided you with feedback or mentorship. The platinum standard for an expert is someone who has cited...

...that can be combined with your gRNA and repair templates of choice though modular cloning steps. The Voytas...

...al., 2007; plasmids at Addgene). AAV vectors: A platform for delivering neuronal tracers AAV vectors have...

..., S.-F., Ravindra Kumar, S., Arokiaraj, C. M., Plattner, V. M., Campos, L. J., Mich, J. K., Monet, D.,...

... the edit should be made HDR requires a repair template and displays reduced efficiency compared to NHEJ...

...High-efficiency blue-shifted channelrhodopsin from Platymonas subcordiformis 445 CoChR Channelrhodopsin from...

... an automated ELISA plate washer. Seal the plate. Place the plate on a microplate shaker set at 400 rpm...use an automated ELISA plate washer. Seal the plate Place the plate on a microplate shaker set at 400 rpm... the plate with a plate seal and wrap in foil. Incubate plate at room temperature on a microplate shaker...each row of a 96-well ELISA microplate. Cover the plate with a 96-well plate seal and incubate at 37 °C... to each well. Cover the plate with a plate seal and incubate on a microplate shaker set to 400 rpm for...

...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...

...into each well of a 96-well plate. By doing this, you are seeding the plate at an average density of 0.5...scan the plate for individual cells, which should be on the same focal plane. Scan the entire plate and make...the 96-well plate, use a microscope to ensure that the cells have detached from the plate and can be isolated...days. Single cells were then plated in individual wells of a 96-well plate and left undisturbed for 13 ...details) Day 1: Seed individual cells in a 96-well plate Day 2–14: Monitor cells for growth and expand cells...tissue culture treated dish, Corning 3516 96-well plate, Corning 3596 0.45 μm polyethersulfone filter, Nalgene... and the specific conditions used. Each 96-well plate described in this protocol could, in theory, give...

...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...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...the Serial Dilution Place a 48-well dilution plate in a chilled 96-well freezer block and place in the...

... 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...tubes Sterile spreading device Reagents LB agar plate (with appropriate antibiotic) LB or SOC media Competent... on ice (approximately 20-30 mins). Remove agar plates (containing the appropriate antibiotic ) from storage...

...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...Bio-Rad, DX200 Thermal Cycler, Bio-Rad, T100 PCR Plate Sealer, Bio-Rad, PX1 1–10 µL single channel pipette...PCR tubes, Axygen, PCR-02-FCP-C ddPCR 96-well PCR plates, Bio-Rad, 12001925 Pierceable foil heat seal, Bio-Rad...µL of each viral dilution to a well of a 6-well plate (1 dilution per well). Leave one well untransduced...

...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...

...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...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...range to anneal to each of the single stranded DNA template strands. The DNA polymerase is also stable enough...

...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...

...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...Particles This protocol is for transfection in a 6 cm plate. The protocol can be scaled to produce different...FBS with penicillin/streptomycin, cells should be plated at this step in DMEM + 10% FBS without antibiotics... you do not want to dislodge the cells from the plate. i. Incubate cells at 37°C, 5% CO 2 for 12-15 hours...