...Autoclavable flasks Sterile pipettes Ice bucket to hold antibiotic Antibiotic at 1000 x concentration disolved...number of plates you’d like to pour. For example: Because we’d like to make 20 plates, and our plates can hold... and negative test results. Sample Data In all cases below (-) indicates that the tested strain is not... Protocols Pouring LB Agar Plates Pouring LB Agar Plates You may...Sterile H 2 O Sterile plates of your desired size - we usually use 60 mm x 15 mm plates which can hold 5-10...colonies, we recommend using larger plates. Many labs use 100 mm x 15 mm plates which can hold 10 - 15 mL of ...appropriate number of plates and stack them on your lab bench. Label the plates with the date and the ...

...middle is hundreds, and the third is tens. Therefore, 1000 µL would read as 100 from top to bottom (as shown... single channel pipettes. These come in standard sizes: P2, P10, P20, P200, and P1000. The table shows...display on these pipettes will also have small tick marks at the bottom. For the P1000, this represents...video for tips on pipetting in the lab. Equipment Pipettes Pipette tips Waste container Containers to hold...Anatomy of a Pipette This front view photo of a P1000 pipette shows the plunger at the top, which is used...tip off of the pipette. This side view photo of a P1000 pipette shows the volume adjustment ring right below...Tips Although there are many different types of pipettes that have different mechanisms to disperse liquids...

...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Ice bucket CO 2 incubator...lots of FBS can promote or inhibit transfection. Test a variety of brands and lots of FBS to find one ...solution through a 0.22 μm filter to remove any precipitates. Stain each well with 1 mL of 0.1% crystal violet...

...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Ice bucket CO 2 incubator...transiently-transfected cells, increases reproducibility, as it eliminates the variation associated with repeated transient...and FBS lots can promote or inhibit transfection. Test a variety of brands and FBS lot numbers to find ... expanded, prepare cell stocks and/or harvest to test for protein expression. Typically, cells transduced...levels of the transgene may have reduced growth rates, especially if that transgene is toxic. Eventually...

...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette 1–10 µL multichannel pipette...dilution is appropriate). Quality of duplicates: Exclude duplicates from analyses if there is more than...-log of the expected titer of the samples being tested. Always include a No Template Control (NTC), i....small aliquot of each standard (enough for 1 or 2 plates) and store at -20 °C. Once a standard is thawed...the assay multiple plasmids containing ITR were tested. Plasmid #59462 is one plasmid that gave reliable...consistent results. Use the recommended plasmid, or test multiple plasmids to find a suitable one. Some labs...95 uL 20X 200X Dilution 3 20 uL Dil. 2 80 uL 5X 1000X Dilution 4 20 uL Dil. 3 80 uL 5X 5000X Dilution ...

...MaxiPreps (Qiagen HiSpeed Max, Catalog #12663) Tips (1000 µL, 200 µL, 10 µL) Bacti Cell Spreaders (VWR, 60828... bioassay plates and remove bacteria. Use one scraper for all plates. Use two 10 mL pipettes (one for ...Antibiotic 245 mm bioassay plates (Molecular Devices, X602) Pro-Tip Pour these plates at least one day in advance...Agar + Antibiotic plates. Prewarm 12 mL recovery media at 37 °C (for at least 15 minutes). Prewarm 3X LB... Agar + Antibiotic plates at 37 °C. Prewarm 8X LB Agar + Antibiotic Bioassay plates. Prechill Micropulser...Micropulser cuvettes on ice. Thaw 4 tubes of electrocompetent cells on ice for 15-20 minutes or until completely...transformed cells on each of the eight bioassay plates (two plates per tube). Distribute evenly with a sterile...

...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette 2–50 µL multichannel pipette...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...

..._CTD_del321-343 TARDBP His T7 ALS Nicolas Fawzi 100092 pET28-his-3C_Tau352 MAPT His T7 Parkinson's, FTD... PRKN mTurquoise2 CMV Parkinson's Pablo Rivera-Fuentes 111723 pBacMam2-DiEx-LIC-N-flag_huntingtin_full-length_Q23...

...Gene and Genome Editing , 3–4 , 100013. DOI: 10.1016/j.ggedit.2022.100013 Zetsche, B., Gootenberg, J. S....interesting adaptive immune response that detects and eliminates foreign DNA, which scientists have modified for... used for small precision edits. NHEJ directly ligates the break ends without the need for a homologous...large genomic deletions or inversions with two gRNA sites on the same chromosome. Most multiplex systems enable...most gRNA targeting sequences will have additional sites of partial homology throughout the genome, called...Directed Repair (HDR) Figure 4: Overview of HDR HDR generates specific genomic edits by copying from a DNA donor...mutation, while whole plasmid or linear dsDNA templates work better for large edits or insertions. When...

...Isoamyl alcohol (49:1) 75% Ethanol RNase-free water or TE solution RNase free tubes: microcentrifuge tubes,...residual ethanol wash and add RNase-free water or TE as soon as the entire tube is dried but while the...visible. Resuspend RNA pellet in RNase-free water or TE. Quantify and assess the quality of your RNA sample...Incubate sample(s) for 15 minutes on ice and centrifuge the sample(s) for 15 minutes at 12,000 × g at 4°C ...Incubate the sample(s) for 2-3 minutes on ice and centrifuge for 15 minutes at 12,000 × g at 4°C to separate...

...cm diameter LB/agar plates without antibiotic High concentration (100 mg/mL, 1000x) carbenicillin stock... fits the plate. Incubate plates at 37 ℃ for 18 hours. Day 2 Observe plates for colony formation. Shown...Antibiotic Plating You may also like... Making LB Agar Plates Bacterial Transformation Recovering Plasmid DNA... the plate at room temperature for at least 30 minutes with the lid on to give the antibiotic time to ... with smaller size than the 1 mg/mL and 2 mg/mL plates and effective selection. Selection Curve of Transformed...

...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...bring up the volume with formulation buffer. Use a P1000 to the bottom of the filter and pipette up/down ...