...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Ice bucket CO 2 incubator...solution through a 0.22 μm membrane. Aliquot 500–1000 μL into sterile tubes. Store the tubes at -20 °C...PEI such that the ratio of μg DNA:μg PEI is 1:3 (1000 μL total per 10 cm dish). Using transfer plasmid...Envelope plasmid, and Transfer plasmid encoding your insert of interest Reagent Preparation DMEM Complete (... optimized for each transfer plasmid. Pro-Tip Endotoxins can inhibit transfection; therefore, plasmid ...

...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Hemocytometer or cell ...system, Nalgene, 565-0010 (or for larger volumes 1000 mL 0.45 µm Rapid-Flow PES Filtration Unit, Nalgene...solution through a 0.22 μm membrane. Aliquot 500–1000 μL into sterile tubes. Store the tubes at -80 °C... deionized water and adjust to a final volume of 1000 mL. Stir at RT until fully dissolved. Pro-Tip This...For each T-175 flask: Aspirate culture media and rinse once with 10 mL of PBS. Aspirate PBS and add 2 mL... at ~80% confluence. Aspirate culture media and rinse once with 60 mL PBS. Aspirate PBS and add 35 mL ... then transfer the cells into a sterile bottle. Rinse the CS2 with 100 mL of DMEM complete medium and ...

...mL conical collection tube. Centrifuge column at 1000 x g for 2 min to remove storage solution. Place ...container. Add 5 mL PBS to the column. Centrifuge at 1000 x g for 2 min to remove PBS. Repeat Steps 30–31 ...resin bed to fully absorb the sample. Centrifuge at 1000 x g for 2 min to collect the sample. Discard column...should be properly disposed of following your institution’s hazardous waste procedures. Last Update: February...February 7, 2022 Protocol Video Watch this instructional video to learn how to affinity purify recombinant... Adjust pH to 7.0 Autoclave or filter sterilize 1000X protease inhibitor cocktail 25 mg leupeptin 50 mg...for your columns, refer to the manufacturer’s instructions. Attach the Gravitrap column to a clamp on a...

...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette CO 2 incubator Pipet controller...96-well plate, seed one of the corner wells with ~1000 cells. This will help you focus the microscope later... incubating the cells. Pro-Tip Use the well with 1000 cells to focus the microscope, then you can easily...cloning cylinders. Workflow Timeline This protocol begins with a stable cell pool. For details, see our protocol...homogenized cell solution to 990 µL complete medium. Instead of transferring 0.125 µL, now transfer 100 times...

...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Ice bucket CO 2 incubator...solution through a 0.22 µm membrane. Aliquot 500–1000 µL into sterile tubes. Store the tubes at -80 °C...µg of DNA into 500 µL of Opti-Pro SFM. Pro-Tip Endotoxins can inhibit transfection; therefore, plasmid ...

...filter, VWR 431229 0.22 µm PES filtering system, 1000 mL, VWR 431098 0.45 µm PES filtering system, 500... aliquots and store at -20 °C until use. BCD TFX 1000 mL BalanCD HEK293 Media 10 mL 10% Pluronic-F68 40...ensure that you are in compliance with your institution’s biosafety regulations. Last Update: February... °C. See below for other Reagent Preparation instructions. Wipe down all pipettes and equipment with 10%...suggest preparing fresh solutions after one month. 1000X protease inhibitor cocktail 25 mg Leupeptin 50 mg...

...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Pipette controller Pipette...your cell pellet but will generally be between 250–1000 µL . The ideal lysis buffer will vary depending ...weight proteins and higher percentage acrylamide when immunoblotting low molecular weight proteins. This...Transfer for 5–6 min for proteins <30 kDa. Transfer for 8–10 min for proteins >150 kDa. Select Start Run...Introduction Western blot is a technique used to separate proteins by size followed by detection using antibodies...Incubate with secondary antibody Video Watch this instructional video to learn how to use western blotting to... Before Starting Refer to the manufacturer's instructions for additional information specific to your ...

...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Pipette controller Microcentrifuge...Pipette tips, 200 µL, VWR 76322-150 Pipette tips, 1000 µL, VWR 76322-154 Pipettes, 5 mL, VWR 89130-896 ...1 or later: Image analysis Video Watch this instructional video to learn how to determine purity and concentration...microcentrifuge. Remove the gel from the plastic wrapper and rinse with deionized water. Gently remove the white sticker...and secure it. Note: Check the manufacturer’s instructions if you are unsure of the correct orientation...the gel. Carefully remove the comb from the gel. Rinse each well with 200 µL 1X MOPS running buffer. Fill... a plastic tray with 100 mL of deionized water. Rinse gel with deionized water for 5 min with gentle agitation...

...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette 20–200 µL multichannel...Microcentrifuge tubes, Neptune 3745.X Pipette tips, 1000 µL, VWR 76322-154 Pipette tips, 10 µL, VWR 76322...protocol describes how to run an indirect ELISA against a purified antigen to demonstrate the expected ...indirect Enzyme-Linked ImmunoSorbent Assay (ELISA) against a purified antigen to validate an antibody. This... the antigen and how to probe with an antibody against the target to show a dose response. Sharing speeds...several days by incubating certain steps at 4 °C instead of room temperature or 37 °C. The protocol notes...minutes. Absorbance was read at 450 nm and plotted against the micrograms of antigen loaded. Last reviewed...

...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette 1–10 µL multichannel pipette...calculate the titer for each sample: $$T = {R*C*D*1000\over V}$$ Where: T = Titer, GC/mL R = Reaction volume...depending on the insert. Please ensure that you are in compliance with your institution’s biosafety regulations... To ensure that the droplets are not disrupted, insert the pipette tips directly in the center of the ...Pro-Tip To ensure that the droplets are not disrupted insert the pipette tips and gently touch the bottom of...

...when using highly competent cells. If you used 100-1000 ng of total DNA in a ligation you will often get...competent cells in your hand instead of on ice Reduce step 4 from 20 - 30 mins to 2 mins on ice before heat-shock...many genetic modifications to create bacterial strains that can be more easily transformed and that will...efficiency, we recommend that you follow the instructions that came with your competent cells. Pro-Tips...out of -80°C and thaw on ice (approximately 20-30 mins). Remove agar plates (containing the appropriate...be approximately 10-fold lower for ligation of inserts to vectors than for an intact control plasmid. ... the competent cell/DNA mixture on ice for 20-30 mins. Heat shock each transformation tube by placing ...

...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Ice bucket CO 2 incubator...ensure that you are in compliance with your institution’s biosafety regulations. Workflow Timeline Day...

...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Reagents 1X PBS Microcentrifuge...staining cells in culture with a primary antibody against a target protein and a fluorescent secondary antibody...labeling cells in culture with a primary antibody against a target protein and a fluorescent secondary antibody...need to be optimized for different cells, target proteins, etc. Sharing speeds science. We believe that ... Before Starting Refer to the manufacturer's instructions for additional information specific to your ...Paraformaldehyde fixation works well for many target proteins, it may not be the best fixation method for all...Remove the paraformaldehyde and follow your institution's laboratory safety guidelines for disposing of...

...middle is hundreds, and the third is tens. Therefore, 1000 µL would read as 100 from top to bottom (as shown...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...come in standard sizes: P2, P10, P20, P200, and P1000. The table shows the pipette with the appropriate...This image shows the P2, P10, P20, P100, P200, and P1000 as shown from left to right. Pipette Dispense Volume... 2 to 20 µL P100 10 to 100 µL P200 20 to 200 µL P1000 100 to 1,000 µL Each pipette should be used with... dispense the correct amount of liquid reagent. P1000: The first red digit is thousands of µL, the middle...

...pipettes Ice bucket to hold antibiotic Antibiotic at 1000 x concentration disolved in the appropriate liquid...use. Negative Result 1: Both Strains Grow Assuming the appropriate strains were streaked on the appropriate...your strains are no longer viable. You can check for this possibility by streaking out both strains on ...antibiotic to the molten gel mix, you should create a 1000x stock solution. For example: If you’ll be preparing...slightly. Even so, you should always use thermally insulated gloves when removing anything from the autoclave...appropriate plates, then if both strains grow, it’s possible that: The antibiotic broke down. You forgot to...concentration for selection. Negative Result 2: Neither Strains Grows If neither strain grows, it's possible that...

...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette 1–10 µL multichannel pipette...titers were similar to those reported by these institutions. This protocol is for a 96-well plate with 20...run: 1.5 h Data analysis: 30 min Equipment qPCR instrument Heating plate Pipettors 1–10 µL single channel...Pro-Tips Use a "Universal" SYBR master mix which contains a high-quality DNA polymerase and a blend of dTTP...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 ...the final volume (mix with >50 uL if your well contains 100 uL) Use a multichannel pipette to load the...tube. Run the following protocol in your qPCR instrument using SYBR detection: 98 °C 3 min / 98 °C 15 ...

...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette 2–50 µL multichannel pipette...modified for other targets. Primers and a probe against the cellular ribonuclease P/MRP 30 kDa subunit ...ensure that you are in compliance with your institution’s biosafety regulations. Last Update: July 7, ...well untransduced (add 150 µL of DMEM complete instead of a viral dilution). Remember, this is an additional... to the GeneJet Genomic DNA Purification Kit instructions. Determine the concentration of each sample ... To ensure that the droplets are not disrupted, insert the pipette tips directly in the center of the ... To ensure that the droplets are not disrupted, insert the pipette tips and gently touch the bottom of...

... or more from the same tube, but unlike using a p1000 tip it will give you more control of where you’re...LiCl selectively precipitates RNA versus DNA or proteins. Add the correct amount of 7.5 M LiCl solution...downstream applications. To improve yield of RNA, instead of incubating at -20°C for 1 hour, you can try ...

...purification and isolation of cells, organelles, lipoproteins, and macromolecules. Importantly, iodixanol ...bring up the volume with formulation buffer. Use a P1000 to the bottom of the filter and pipette up/down ...of contaminants in each fraction. * AAV capsid proteins VP1, VP2, VP3; M protein marker. Image adapted...