...video content, including how-to screen captures, lab procedure protocols, and career advice videos....guides for standard laboratory procedures. How-To Videos Screencapture walk-throughs to help guide you...Inoculating a Liquid Bacterial Culture Protocol Getting Started with Tissue Culture Tips and tricks for working...working with tissue culture in the lab. Blog post: 10 Basic tips for mammalian cell culture Aseptic Technique...Related Page Making LB Agar Plates Create plates to culture bacteria in the lab Pouring LB Agar Plates Protocol... Over-Agar Antibiotic Plating Create plates to culture bacteria in the lab Over-Agar Antibiotic Plating...Transformation Protocol Inoculating a Liquid Bacterial Culture How to prepare and grow bacteria in a liquid medium...

...the cartridge with the DG8 gasket, making sure that it is secure. Transfer the cartridge holder to the droplet...is not at temperature, touch the screen on the plate sealer to allow it to get to temperature. Once the...generally considered biosafety level 2+. Please ensure that you are in compliance with your institution...Purification Kit, Thermo Fisher, K0721 6-well tissue culture treated dish, VWR, 29442-042 DMEM, high glucose...15 mL DMEM Complete 3 µL of 250 U/µL benzonase Procedure Transducing Cells Thaw the virus on ice. Prepare...Gently aspirate media from the 6-well plates. Be sure to use a new aspirating pipette for each well as... it in the Biological Safety Cabinet (BSC). Make sure that the BSC is supplied with sufficient pipette...

...PCR machine. Set annealing temperature 5°C below the primer melting temperature (Tm). Set extension step... See manufacturer’s instructions for specific instructions about extension time and temperatures. Initial...designed to have similar melting temperatures (Tm). Set the annealing temperature to 5°C lower than the Tm of...radical temperature changes during a typical PCR. The DNA polymerase has an optimum temperature around ...withstand many heating and cooling cycles, which would denature DNA polymerases from other species. In addition...denaturation step is commonly performed at 94–98°C. Denature 30 seconds at 94°C: Continued denaturation of ...forward and reverse primers are stable within this temperature range to anneal to each of the single stranded...

...puncturing your glove. Ensure that the tip is properly set on the pipette to ensure pipetting accuracy. ...working in a laboratory, properly dispensing liquid ensures the accuracy of your experiment and any changes...Pipette tips Waste container Containers to hold measured liquid (ex: microfuge tube, bottle, etc.) Labels...read as 100 from top to bottom (as shown in the picture above), while 650 µL would read as 065. P200: The...read as 100 from top to bottom (as shown in the picture above), while 95 µL would read as 095. P20: The...Therefore, 10 µL would read as 100 (as shown in the picture above), while 2.2 µL would read as 022. P2: The...Therefore, 1 µL would read as 100 (as shown in the picture above), while 0.5 µL would read as 050. Each volume...

...electrophoresis chamber and secure it. Note: Check the manufacturer’s instructions if you are unsure of the correct...follows: Figure 2 Using the box tool, draw a box around the entire first gel lane (as in Figure 2). Select...with Coomassie blue, protein band intensities are measured using ImageJ software and a standard curve is ... mL of deionized water. Gently invert to mix. Procedure Section 1: SDS-PAGE Add 5 µL of 4X sample buffer...Fill the chamber with 1X MOPS running buffer. Make sure that the chamber is sealed. Load 5 µL of the prestained... of the gel with an appropriate imaging system. Figure 1 Recombinant antibody preps should have 2 clear...electrophoretic mobility of the bands (sample AR0016 in Figure 1) indicates that the samples may not have been...

... web-based primer design software to ensure a melting temperature between 50-60°C for your PCR primers...room temperature, then add 1 μl of 25 mM EDTA, followed by another 5 minutes at room temperature. The ...separation of linearized vector from the reaction mixture, and may provide counter-selection for negative...negative clones as in the example shown here. The manufacturer of an LIC-specific vector will provide the homologous...LIC is often as simple as using the backbone manufacturer's suggested leader sequence fused to your gene...separate the linearized vector from the reaction mixture by gel electrophoresis followed by gel purification...products with sterile water (instead of TE buffer) to ensure optimal salt concentrations in subsequent reactions...

...These lab safety guidelines provide steps to ensure you are working in BSL-1 and BSL-2 labs safely....different types of hazards and to provide protective measures for each level, as each level has different safety...pathogens including organisms such as Staphylococcus aureus or Vibrio cholerae . BSL-2 includes all of the ...precautions to prevent injuries, ingestion, and exposures to hazardous materials. Video Watch the video ...laboratory. Before you start your experiment, make sure your workspace is clean and uncluttered. Disinfect...the appropriate contact times for each agent. Make sure you have enough space to work at your lab bench....are also required to be present in the room. Make sure that you know where these are located before you...

... some cells in the culture may adversely affect the surviving cells in the culture, so it is important...blasticidin), which enables selection of a stable cell culture after transduction. Performing antibiotic selection...Microcentrifuge tubes, Neptune 3745.X 6-well tissue culture treated dish, Corning 3516 15 mL conical tubes,... be subjected to multiple freeze-thaw cycles. Procedure Before beginning, determine the optimal dose of... mL of virus solutions at various dilution. Make sure to use the polybrene-containing media to make the...stable cell pool. Observe the dish every day to ensure that the cells in the untransduced well (0 µL lentivirus... reduce the concentration of the antibiotic in culture, or remove the antibiotic entirely. If the antibiotic...

...buffer (see manufacturer's instructions) Gel loading dye Electrophoresis buffer Pipet tips Procedure Select...at appropriate temperature (usually 37 °C) for 1 hour. Always follow the manufacturer’s instructions. .... Follow the manufacturer’s instructions. If your enzyme did not cut, check to make sure that it isn't... like... Recovering Plasmid DNA from Bacterial Culture Purifying DNA from an Agarose Gel DNA Ligation ...amounts) Appropriate restriction enzyme (see manufacturer's instructions for proper ammount) Approrpriate...Restriction Enzyme(s) Buffer BSA (if recommended by manufacturer) dH 2 O up to total volume Pro-Tips The amount...window) QIAGEN DNA cleanup kit . See the enzyme manufacturer's instructions for more details. To visualize...

... at -20 °C Procedure Section 1: Affinity chromatography Equilibrate filtered tissue culture supernatant...affinity purify recombinant antibodies from cell culture supernatant using Protein A or Protein G columns... following your institution’s hazardous waste procedures. Last Update: February 7, 2022 Protocol Video..., binding buffer, and elution buffer to room temperature (RT) before use. Reagent Preparation 1 M sodium... containing the recombinant antibody to room temperature (see Transfection for Recombinant Antibody protocol...part Protein A/G binding buffer to 1 part tissue culture supernatant and mix well. Uncap the Protein G Gravitrap...