...transduction of the trabecular meshwork following intracameral injection in mice, rats, and dogs. This serotype...

...with an empty tube. Alternatively, you can just subtract the weight of the empty tube from the weight of...the (Link opens in a new window) QIAquick Gel Extraction Kit . Always follow the manufacturer's instructions...

... glass bottle: 4 g NaCl 4 g Tryptone 2 g Yeast Extract and dH 2 O to 400 mL Note: If your lab has pre-...mL Kanamycin 50 µg/mL Spectinomycin 50 µg/mL Tetracycline 10 µg/mL Tips and FAQ What is the difference...

...experimental procedures, such as minipreps and RNA extractions. Plus, centrifuges are useful for simply collecting...cousins, but can hold much larger containers. Ultracentrifuges are also generally floor sized centrifuges...

...Community at Protocols.io Addgene's Protocol for RNA Extraction Without A Kit References ACE2 - Andersen, KG,...

... g pre-mixed powder consisting of: 5.0 g yeast extract 10.0 g peptone from casein 10.0 g sodium chloride.../mL 50 µg/mL Spectinomycin 50 mg/mL 50 µg/mL Tetracycline 10 mg/mL 10 µg/mL Notes: Unless otherwise indicated...

... a sonication step may be required for hard to extract proteins such as those in the nucleus. Incubate...absorbance of the duplicate samples on the plate. Subtract the average absorbance of the blank from all of...

... min at 100 x g . Gently aspirate supernatant. Extract genomic DNA according to the GeneJet Genomic DNA...used to transduce HEK293T cells. Genomic DNA is extracted from the target cells and assayed for integrated...

...non-integrating episomes to allow sorting and dosage tracking of reprogramming factors Fluorescent tagged episomals...

...using various delivery methods including intracerebroventricular (ICV) delivery in newborns, and retro-orbital...

...yes, activate S. pyogenes Marraffini pSimpleII-U6-tracr-U6-BsmBI-NLS-NmCas9-HA-NLS(s) 47868 Mammalian U6...

...Pro-Tip When you weigh out a reagent, you may leave trace amounts of it on the balance. Therefore, it’s best...

...cells AAV Purification by Iodixanol Gradient Ultracentrifugation Protocol Mulitchannel Pipetting Technique...

...again but store at 4 °C and use within 1 week. Keep track of the Ct value for each standard over time. They...

...Anti-Cas9 Western blotting was performed on whole-cell extracts from the initial stable cell pool (pool) and from...

... milk going to help me extract DNA from agarose gel?!”   The DNA gel extraction with glass milk method... a similar workflow as a column-based plasmid extraction kit. First, you lyse the bacteria and denature...contamination and you can use a phenol chloroform extraction to remove remaining proteins and RNase. Finally...ready for use! Plasmid purification and DNA gel extraction with glass syringe filters Starting material:...fragments The plasmid purification and DNA gel extraction method described in Kim and Morrison is nearly...protocol and guanidine isothiocyanate for the gel extraction protocol) disrupts the DNA’s hydrogen bonds with... more flexible than commercial kits.  DNA gel extraction with glass milk Starting material: agarose gel...

...information from a region (anterograde tracer, Fig. 1B). Tracers are typically delivered to a particular... either trace downstream from the soma (neuron cell body) toward the axon (anterograde tracer) or upstream...making them useful tracers, but finding a virus that works well for retrograde tracing can be difficult....brain works. There are two methods of neuronal tracing (Fig. 1): one to visualize neurons that send information...information to a region (retrograde tracer, Fig. 1A), and another to visualize neurons that receive information...from the axon terminals to the soma (retrograde tracer) (Fig. 2). Viruses as neuronal scalpels Viruses...transport and can therefore perform retrograde tracing, but often at the cost of toxicity, low expression...

...days. The crRNA and tracrRNA are annealed on a PCR block and the annealed cr:tracrRNA is then mixed with...Anneal crRNA and tracrRNA using a PCR block. Add Cas9 protein to the annealed cr:tracrRNA and incubate at...Tag 96 genes at a time using Custom Synthetic cr:tracrRNA/Cas9 Ribonucleoprotein (csRNP) delivery Epitope...started off by using custom synthetic two-part cr/tracrRNAs for CRISPR experiments (in collaboration with ...-tagging experiments. Cas9 protein, annealed cr:tracrRNA, and single-stranded donor DNA. The success of...have recently demonstrated that the two-part cr:tracrRNA system is much more potent than the in vitro-...Dewari et al. 2018 and Figure 2). Not only is the cr:tracrRNA approach highly efficient, it is also straightforward...

... challenging to track intracellularly, but aptamers permit the study of intracellular RNA dynamics. Aptamers...its ability to track RNA dynamics in a model of Fragile-X tremor/ataxia syndrome (Strack et al., 2013)....target cellular RNA to specific intracellular locations using Cas9 and track the localization of the complex... molecule. Fluorescent RNA aptamers are used to track cellular processes by transforming cells with plasmids...then, Spinach2 has been used to investigate intracellular viral DNA dynamics in neuronal cells (Nilaratanakul...is better able to maintain its fluorescence intracellularly due to a lower dependency on magnesium and ...bright, high-affinity complex for RNA labeling and tracking." ACS chemical biology 9.10 (2014): 2412-2420....

...pre-amplified stock. A last option is to perform a gel extraction of either the original sample or the amplified...