...workshop was well received, and then when the team started brainstorming ideas for the next workshop, they...

...method to create CRISPR/Cas9 constructs, assemble standard plasmids parts in different combinations, or other...

...organism where deletion strains are available. Then, staff or students clone similar sequences from a less-...

...2020 by Jennifer Tsang. CRISPR-Cas technology is constantly evolving. Variants of Cas proteins can be used...

...away from a desired site does not have the same devastating impact on efficiency as 10-15 bps away does. ...

...In order to understand the complex wiring of the brain and the underpinnings of neurological disease,...

...aa 155), includes I152L in N-terminal fragment Constant rate of p53 tetramerization in response to DNA...

...where cleavage leads to the loss of spectinomycin resistance. xCas9 3.7 showed the broadest PAM range, recognizing...

...useful tools, and as such, we’re always looking to understand our community better. We therefore looked at ...

...translate perceived signals to physical actions. Understanding neuronal pathways and the function of certain...

...techniques do tip the scales in your favor. For starters, Golden Gate cloning is extremely efficient due...

...not a leader,” because you don’t see yourself  standing in front of a crowd and using your charisma to...

...and adaptations that can provide an increased understanding of DNA binding and gene regulation. For additional...

...extremely important gene in animal development. To understand where sonic hedgehog got its name, we need to...

...medical journal. Decades later and after years of painstaking work trying to isolate the causative gene for...

...IgG isotype standard 2.5 mg/mL - can use commercial standard or validate an in-house standard Before Starting...Thaw IgG standard and prestained protein ladder on ice. Reagent Preparation Prepare IgG Standard Dilute ...ready-to-use bio-safe Coomassie G-250 stain (Addgene uses SimplyBlue SafeStain) and ImageJ software. The sample... serial dilutions of a standard antibody of known concentration. After staining with Coomassie blue, protein...ready-to-use biosafe Coomassie stain and ImageJ software. Protocols... Protocols Coomassie Purity Stain of Recombinant Antibodies...Antibodies Coomassie Purity Stain of Recombinant Antibodies You may also like... Antibody Plasmid Collection...

... Protocols Generating Stable Cell Lines with Lentivirus Generating Stable Cell Lines with Lentivirus... antibiotic resistance (e.g., puromycin, blasticidin), which enables selection of a stable cell culture...lentiviral vectors deliver antibiotic resistance. This protocol was established using 293T cells but can be adapted... of antibiotic, which may not be stable at 37 °C. To achieve a stable cell pool, the antibiotic selection...expression. As polyclonal populations of resistant cells start proliferating and the individual wells become...Protocol to generate stable cell lines expressing a gene of interest from an integrated lentiviral vector...Introduction This protocol can be used to generate stable cell lines expressing a gene of interest from an...

...help stabilize the standards add carrier DNA to a final concentration of 4 ug/mL to each standard dilution... Once a validated standard curve is obtained, make a small aliquot of each standard (enough for 1 or 2...their initial value. If the Ct value of the standard starts to drift, it’s time to make a new one. When...your plasmid standard with DNase ** Make 6 serial dilutions, in duplicate, of your standard curve plasmid...5'-CGGCCTCAGTGAGCGA ITR-containing plasmid for standard curve RNase-free DNase 10X DNase buffer Nuclease-free...Pipette tips General Considerations Always run standards and samples in duplicate at least If possible,... the samples have been added to the qPCR plate. Start by adding water, then SYBR master mix, then the ...

... carry one or more antibiotic resistance genes, which confer resistance to a specific antibiotic to the...to be resistant to the antibiotic. On the second plate, streak out a strain that’s not resistant to the... be resistant to the antibiotic, (+) indicates that the tested strain is supposed to be resistant to the...Negative Result 3: Only the Non-resistant Strain Grows If only the non-resistant strain grows, it’s likely ... carrying them. The presence of an antibiotic resistance gene on a plasmid allows researchers to easily...for the selection of only those bacteria with resistance to that antibiotic - usually conferred by a plasmid... plasmid carrying the antibiotic resistance gene. The following protocol will allow you to make your own...

...transfer stack. Set the Top Stack to one side and discard the white separator. Keep the Bottom Stack in the...SDS-PAGE running buffer Prestained protein ladder Ethanol iBlot 2 PVDF Mini Stack, Thermo Fisher IB24002... of the BSA standard that range from 0–2000 µg/mL . In duplicate, dilute 10 µL of standard, blank, and...blank from all of the samples. Plot a standard curve of BSA standard concentration versus absorbance. Extrapolate...antibody Secondary antibody Deionized water Before Starting Refer to the manufacturer's instructions for additional...total protein concentration of the sample from the standard curve. Determine the volume of sample required...the samples into the gel. Load 5–10 µL of the prestained protein ladder. Place the lid on the tank and...