...lens which magnifies the image, then the ocular lens, where the image is further magnified before it finally...4x objective magnifies the image four times the actual size, the 10x objective magnifies it 10 times, ...also 10x, you would actually be seeing a 100x magnified image of that object! Equipment Light Microscope...aperture are in focus. Center the aperture in your field of view and then slowly open the diaphragm until...the stage (and ultimately your slide) until your field of view is centered on a region of interest. If ...

...necessary to remove iodixanol prior to use of your purified virus. Nonetheless, we recommend performing a ...a buffer exchange before using the purified AAV in vivo . The iodixanol gradient in this protocol is composed...genome containing) particles. On average, iodixanol-purified AAV preparations contain ~20% empty particles....60% iodixanol step Carefully add up to 5 mL of clarified supernatant on top of the gradient. Use 1X PBS...fractions obtained from the 40% phase contain the purified AAV. Option #3 Puncture the QuickSeal tube slightly...vertical black line indicates the location of the purified AAV. Left image adapted from Zolotukhin, S., et...the position of the needle for harvesting of the purified AAV using option #1. Right image adapted by permission...

...Microsoft Excel or similar software Reagents Purified antigen Purified recombinant antibody Isotype control antibody...describes how to run an indirect ELISA against a purified antigen to demonstrate the expected antibody dose...Enzyme-Linked ImmunoSorbent Assay (ELISA) against a purified antigen to validate an antibody. This protocol...the Antigen Standard and coat the plate Dilute purified antigen to 20 ng/µL in a microfuge tube and vortex...concentration. Prepare serial dilutions of the purified antigen as follows: 2 ng/µL : Add 100 µL of 20...coated with serial dilutions of human recombinant purified Desmin (blue) or a negative control protein, Apolipoprotein...

...Many molecular biology techniques require highly purified and concentrated plasmid DNA. This page will discuss...TE. You will now have plasmid DNA that has been purified away from the bacterial proteins and genomic DNA...become sheared and will therefore contaminate your purified plasmid DNA. Incubate solution on ice for 5 min...plasmid DNA. To improve the purity of plasmid DNA purified without a kit it is advisable to perform a phenol...

...of the primer that binds to the sequence to be amplified (usually 18-21bp). When selecting restriction ... primer that hybridizes to the sequence to be amplified (the ORF in this case), not the Tm of the entire...sufficient amount of DNA). Once you have cut out and purified your insert and vector bands away from the gel...diagnostic restriction digest of 100-300ng of your purified DNA with the enzymes you used for the cloning....

...imaging system ImageJ or similar software Reagents Purified recombinant antibody, 0.9–1.1 mg/mL SimplyBlue...the recombinant antibody Dilute or concentrate purified recombinant antibody to a concentration of 0.9...platform. Pour off the water in the sink. Take a brightfield image of the gel with an appropriate imaging ... that have a purity of ≥90% but some labs use unpurified tissue culture supernatant routinely without ...

...protocol to perform digital droplet PCR (ddPCR) for purified AAV titration....how to use droplet digital PCR (ddPCR) to titer purified recombinant Adeno-associated viral vectors (AAV...the ITR elements in the viral vectors but can be modified for other targets. The dilution series outlined...ddPCR for AAV quantification. This protocol was modified from the publication Lock et al. (2014) (Link ...

...restriction enzymes (e.g. BsaI), which cut at a specified distance from their recognition sequence. This... end will now look like this: Note: Elute your purified digestion and PCR products with sterile water ...purification . Step 5: Create Insert Overhangs Treat your purified PCR product with T4 Pol in the presence of dCTP...

...purification. Electrophoresis uses an electrical field to move the negatively charged DNA through an agarose...temperature for 20-30 mins, until it has completely solidified. Pro-Tip If you are in a hurry, the gel will ...well, instead of diffusing in the buffer. Once solidified, place the agarose gel into the gel box (electrophoresis...

...transformation and basic knowledge of the library being amplified. Please see our protocol on Bacterial Transformation... change in representation compared to the pre-amplified stock. A last option is to perform a gel extraction...extraction of either the original sample or the amplified sample, followed by reamplification of the DNA,...

...to a plate after it's already been poured and solidified..... With a 6 cm diameter petri plate containing solidified LB-agar, pipette 150 µL of carbenicillin on top...

...) transfection protocol. This procedure can be modified for alternative packaging cell lines or transfection...without limiting cell growth. Left panels: bright field images; right panels: GFP channel images. Last reviewed...

...and expression, the recombinant antibody can be purified for use in a variety of applications. Sharing ... used as-is or the recombinant antibody can be purified from the supernatant (see our Recombinant Antibody...

...look as you expected. Once you have cut out and purified your insert and recipient plasmid backbone bands...diagnostic restriction digest of 100-300ng of your purified DNA with the enzymes you used for the cloning....

... and downstream (3’) of the DNA segment to be amplified. When these reagents are combined in an appropriate... After the PCR is complete, the product can be verified based on size by gel electrophoresis . Protocol...

...and stack as you pour. If your agar partially solidifies in the bottle while you’re pouring, you should...stored at 4 ℃ until use. Once your plates have solidified and dried, you should test them to make sure ...

...has been optimized for adherent cells but can be modified for suspension cells. The number of monoclonal...the cells are overly confluent or the medium is acidified (i.e., the medium appears orange/yellow if using...

...protocol for DNA quantification , which can be modified for RNA. Store your RNA sample(s) at -80 °C to...protocol for DNA quantification , which can be modified for RNA.Store your RNA sample(s) at -80 °C to ...

...response element (RRE) in the target cells but can be modified for other targets. Primers and a probe against... on their particular sample. This protocol was modified from the publication Wang et al. (2018) . Before...

...restriction enzyme digestion. However, once you have identified the positive clones, it is important to verify...serum-free media Invitrogen: #31985 Dulbecco’s Modified Eagle Medium (DMEM) Invitrogen: #11995 Fetal Bovine...