...Tetracycline 10 mg/mL 10 µg/mL Notes: Unless otherwise indicated, the antibiotic powder can be dissolved in dH ...test results. Sample Data In all cases below (-) indicates that the tested strain is not supposed to be resistant... resistant to the antibiotic, (+) indicates that the tested strain is supposed to be resistant to the ...

...primers produce inaccurate, nonspecific DNA amplification product, and long primers result in a slower...which creates primer dimers and disrupts the amplification process. When designing, if unsure about what...

...1) and biosafety level 2 (BSL-2). The BSL-1 classification is for labs working with low-risk microbes ...choose to reuse or repurpose this SOP in another location, please note that you do so at your own risk; ...

... these cells can still be used in downstream applications. Safety Warnings Lentivirus is generally considered...without calcium or magnesium, Corning 21-040-CV (cations can affect the attachment of adherent cells) 0.45...

... room. Make sure that you know where these are located before you start. Use the eyewash station if unwanted...choose to reuse or repurpose this SOP in another location, please note that you do so at your own risk; ...

...Posts Western Blot Protocol Recombinant Antibody Purification Protocol Molecular Biology Reference Introduction...as methanol or acetone may be better for some applications. Remove the paraformaldehyde and follow your...

... length in base pairs) for visualization and purification. Electrophoresis uses an electrical field to...instructions on how to do this, visit the Gel Purification page. Tips and FAQ How do you get better resolution...

...without calcium or magnesium, Corning 21-040-CV (cations can affect the attachment of adherent cells) Trypsin...orange/yellow if using a standard phenol red pH indicator) because the buildup of waste products could be...

...post on ddPCR for AAV quantification. This protocol was modified from the publication Lock et al. (2014) ...

...plate to inoculate a bacterial culture for DNA purification will minimize the chance of having a mixture...

...in the photo above has a capacity of 200 g, as indicated by the number above the "Zero" button. Before ...

...bring your materials to a particular temperature, catalyze chemical reactions such as restriction digests...

...centrifuge begins to wobble or shake, this could indicate an unbalanced or improperly loaded centrifuge....

...inhibit transfection; therefore, plasmid DNA purification should include an endotoxin removal step. For...

...product, gel-purify DNA segments. Otherwise, PCR purification or even the raw PCR mix can work fine in an ...

...subsequently isolated by affinity purification against the epitope tag. After purification of the locus, molecules...conventional methods for genomic purification, CRISPR-based purification methods are more straightforward...protein through mutation of its catalytic domain. Fusions to catalytically inactive Cas13 can be used to...table above, 3' and 5' indicate on which end of targeted sequence the PAM is located. The majority of the...unintended genomic locations On-target activity Cas9 cleavage at a desired location specified by a gRNA...Resources Web References PAM Sequences Glossary Publications CRISPR Overview Class 2 C lustered R egularly...genes in various cell types and organisms, but modifications to various Cas enzymes have extended CRISPR ...

...Handling Visualization Signal Amplification Application Overview Quantification Methods Capture Methods Visualization...Antibody Applications Antibody-based applications can be generally classed into four categories: Quantification... Read on to learn more about the applications common to each category, including an outline of the basic...these applications, visit the Antibody section of the Addgene Protocols page. Antibody Applications - Quantification...concentrates in the sample. Controls for quantification methods Quantification methods should be run with: a positive...using sonication to break DNA up into fragments of 300-1000 bps in length. Note: This sonication process...production and storage techniques, and explain common applications. Science...

... Origin of Replication ( ori ) DNA sequence which directs initiation of plasmid replication (by bacteria... Educational Resources Molecular Biology Reference Molecular...plasmid selection DNA sequencing for plasmid verification Sanger Sequencing Next Generation Sequencing...Techniques in chemistry enable isolation and purification of cellular components, such as DNA, but practically...double-stranded DNA that typically carry genes and can replicate independently from chromosomal DNA. Although they...the bacterial chromosomal DNA, plasmid DNA is replicated upon cell division, and each daughter cell receives...manipulating genes. For instance, most plasmids are replicated in E. coli and are relatively small (∼3000 - ...

...therapeutic applications. Clinical trials using AAV for various gene delivery applications are currently...adenovirus. These genes (E4, E2a and VA) mediate AAV replication. The transfer plasmid, Rep/Cap, and the helper...integrations may occur at a much lower frequency. As a replication-incompetent virus, AAV cannot enter the lytic...double stranded synthesis. These episomes can concatemerize, producing high molecular weight structures ... are thought to be maintained in episomes or concatemers. Episomes differ profoundly from viral particles...gives a summary of the tropism of AAV serotypes, indicating the optimal serotype(s) for transduction of a...serotypes are denoted using a slash, so that AAV2/5 indicates a virus containing the genome of serotype 2 packaged...

...Channelrhodopsins: cation channels from Chlamydomonas reinhardtii ChR2 Widely used light-gated cation channel from...light of the correct wavelength, the pore opens, cations flow into the cell (yellow dots), and the neuron...reporters of molecular signals; e.g., calcium indicators In this overview, we will focus on the common... electrochemical responses, such as nonspecific cation influx vs. proton efflux. Researchers have used...is excited by blue light and permits nonspecific cation influx into the cell when stimulated. Channelrhodopsin...optogenetic tool, is also a blue light activated cation channel. ChR2 is preferred over ChR1 because ChR2...been expanded by scientists through both the identification of novel ChRs from other algal species and ...