...of a compound light microscope with the main components labeled: Figure 1: Diagram of a compound light...electron microscopes that allow us to see cellular components with science-fiction levels of detail. You can...common types that you're likely to encounter: the compound light microscope. As the name suggests, light ...light to detect small objects. Two of the most important properties of a microscope are magnification (... make it through the sample because different components of the specimen will refract and reflect the ...) using dyes or stains that add color to some components of the sample but not others. The light that ...see several pieces of information, but the most important (for now) is the magnification power, such as ...

...product: Run PCR to amplify your insert DNA. It is important to use a high fidelity taq polymerase to minimize.... The fidelity of the polymerase becomes more important the longer the expected PCR product is. You should...some DNA during the gel purification step, it is important to digest plenty of starting material. We recommend...be cut with both enzymes, and therefore it is important that the digest goes at least 4 hours and as long...running a gel for purification purposes it is important to have nice crisp bands and to have space to ...expected. When cloning by PCR, it is especially important to run the product on a gel. This allows you to...insert and vector bands away from the gel, it is important to determine the concentration of recovered DNA...

...some DNA during the gel purification step, it is important to digest plenty of starting material. We recommend...be cut with both enzymes, and therefore it is important that the digest go at least 4 hours and as long...running a gel for purification purposes it is important to have nice crisp bands and to have space to ... your favorite gel purification method, it is important to determine the concentration of recovered DNA...ratios of recipient plasmid to insert. It is also important to set up negative controls in parallel. For instance...

... and using a centrifuge to separate different components in a liquid sample....laboratory equipment used to separate different components in a liquid sample. It achieves this by using... you use, most will have essentially the same components. The photo to the left shows a front view of ...supernatant, or both. Conclusion Centrifuges are important tools for many different protocols. Improper use...

...around the band as possible. To do so, it is often important to take the excised band, lay it down on the UV...sides with the razor blade. This is especially important during the DNA purification step, as many kits...manufacturer's instructions. Note: It is usually important to determine the concentration of the DNA that...

.... Transformation of bacteria with plasmids is important not only for studies in bacteria but also because...amplification. Higher efficiency cells are more important if you will be transforming with very small amounts...critical for Ampicillin resistance but is much more important for other antibiotic resistances. Plate some or...

...transduction efficiency by facilitating nuclear import of the vector’s preintegration complex in the transduced...below. Do not change the ends; these bases are important for cloning the oligos into the pLKO.1 TRC-cloning...temperature. This will take a few hours, but it is important for the cooling to occur slowly for the oligos...you have identified the positive clones, it is important to verify the insert by conducting a sequencing...

...Background Information Bacterial glycerol stocks are important for long-term storage of plasmids. Although you...completely and will improve the shelf life. It is very important that you shake the glycerol before freezing (5...

...initials. Labeling within a laboratory setting is important for organization, and it is recommended that you...white dot growing on the solid medium. This dot is composed of millions of genetically identical bacteria ...

... Digest Introduction Sequence verification of important plasmid features (such as the gene/insert, fusion...you sequence any portion of the plasmid that is important for your experiments. There are a number of factors...

...Appropriately weighing reagents is important to ensure the accurate composition of your solutions and buffers...

... A typical T4 Pol reaction is shown. Mix the components (add the T4 last) and incubate at room temperature... by your polymerase manufacturer. It is very important to remove all free nucleotides from your PCR product...

... and using a centrifuge to separate different components in a liquid sample Using a Light Microscope Learn...primers Watch the Video! Sequence Analysis Verify important plasmid features using sequence analysis RNA Extraction...

...desired insert. If you are in this situation, it is important to treat the digested vector backbone with a phosphatase...setting up the ligation reaction itself, it is important to determine the amount of cut insert and vector...

...Conclusion Pipetting accurately and precisely is important since even small changes in the volume could affect...Beyond knowing the proper way to pipette, it’s important for scientists to calibrate their pipettes to ...

...organelles, lipoproteins, and macromolecules. Importantly, iodixanol is inert and non-toxic to mammalian...vivo . The iodixanol gradient in this protocol is composed of steps that separate out contaminants from an...

...of cell death upon antibiotic selection. It is important to monitor these cells regularly and replace the... the surviving cells in the culture, so it is important to do regular media changes and maintain optimal...

...intensity of the protein bands to the overall lane. Import the gel image into ImageJ. Select File . Select...in Google Sheets. Open exported CSV file. Select import. Choose file. Format as needed. Select the Insert...

...the pellet for 5-10 minutes. *Critical* It is important to not let the pellet get too dry before resuspending...

... different stages of molecular cloning, it is important to get a quick and accurate reading of DNA concentration...