...lentiviruses easily made in your own lab. We provide protocols and useful tips for generating your own lentivirus...using this plasmid Cas9 expression is typically cytotoxic, which makes generation of high titer Cas9 lentiviruses...lentiviruses challenging. This cytotoxicity also causes growth rate variability when using lentiCas9-blast...

...In 2017, Lenny Teytelman, CEO of protocols.io, organized a panel to discuss reproducibility issues in...2018 and involved presentations and a handout on protocol sharing, material sharing, computational reproducibility...management plans, electronic lab notebooks, wet lab protocols, reagent sharing, bioinformatic tools, data and...

...selecting a bright fluorescent protein and a photostable fluorescent protein. In the third post of this...applications (e.g. labeling of cells or tracking promotor activity), but it is problematic in applications...verify a number of key properties (brightness, photostability, oligomerization) of a number of promising ...

... area. Unless you are looking for a particular protocol or are trying to replicate an experiment, the ...to the methods section for finer points of the protocols. Are there any red flags about how the work was...Louisiana State University. She is a fan of photosynthesis and writes about autotrophs and biochemistry...

...the Type IIS restriction enzyme and thermocycler protocol. It will also identify overhangs with a high potential...possible, NEB also optimized the thermocycler protocols and made sure that T4 ligase was the correct choice...screening for selection, failed under normal cycling protocols, and required a 48-hour incubation at 37 °C. While...

...viruses also have a propensity to integrate into proto-oncogenes, which can lead to cancer. These viruses...reason for lab accidents is not following safety protocols.) If administering virus to mice, part of your... be safe is to know and closely follow safety protocols. This is especially true when you’re tired, in...

...drawbacks of different methods (for comprehensive protocols for all of the methods discussed here refer to...staining. Check Out Our In-Depth Fluoresence Titering Protocol Another popular approach utilizes the antibiotic...pseudotyped HIV-1-based lentiviral vectors."Nature protocols 4.4 (2009): 495-505. PubMed PMID: 19300443. 2....

...CRISPR-Cas activity. For example, fusing AcrA4II to the photo-sensitive LOV2 domain allows anti-CRISPR activity... blue light (Bubeck et al., 2018). 3. Reduce cytotoxicity of CRISPR-Cas editing in tissues The small size... systems prevents the off-target effects and cytotoxicity that result from excessive nuclease activity...

... and delivers it cargo to certain cell types.  Botond Roska’s lab at the Institute of Molecular and Clinical...types (ex: rods or cones) or cell classes (ex: photoreceptors which encompassed both rods and cones).  Targeted...also expressed. If Cre-mCherry expresses in photoreceptors and horizontal cells, and Cre-dependent-GFP...

...Resources on addgene.org Western Blot Protocol Western Blot Protocol Video ...Find other tips and tricks for molecular biology protocols Check out other Antibody 101 articles Resources...

... microbes? Try your hand at photography and participate in a science photo competition. Or nominate yourself...Indonesia. Since then, I have tried many things: photography, making videos, presenting in pubs and at science...

... of a BSL-3 lab. Jesse Bloom’s lab published a protocol and reagents for pseudotyping lentivirus with ...methods to use CRISPR to detect SARS-CoV-2. Other protocols and tools for detection have been made openly ... Pettie D, King NP, Balazs AB, Bloom JD (2020) Protocol and Reagents for Pseudotyping Lentiviral Particles...

...behaviors - from basic types like locomotion and motor coordination to more complex situations such as ... using recombinant adeno-associated virus. Curr Protoc Microbiol. (2013); Chapter 14:Unit14D.5. PubMed...Learn about Addgene's viral service Find viral protocols and videos Find AAVs for your research ...

... process, check out the Addgene AAV Production protocol. Titering Once you’ve produced, harvested, and...process, refer to the Addgene AAV Titration by qPCR Protocol. If you’re interested in the ddPCR titering process...Transfer PlasmidAAV Variables That Matter AAV protocols on addgene.org  AAV Titration by qPCR Using SYBR...

... Find basic molecular biology protocols Find plasmid cloning protocols Check out our molecular biology...Number of fragments assembled Golden Gate Assembly protocol* (Volume of 1mL Outgrowth plated) Correct assemblies...

...figure 1.   Find Additional Useful Protocols on Addgene's Protocol Pages Agarose cloning method procedure...Vectors Resources on Addgene.org Check Out Our Protocols Pages Browse Fluorescent Protein Plasmids Find...

...Resources on Addgene.org Protocol: Polymerase Chain Reaction (PCR) Protocol: How to Design a primer  ...variants based on real-time LightCycler PCR. Curr Protoc Hum Genet. 2014;80:7.21.1-7.21.. Published 2014...

...genome, removing the risk of oncogenicity or genotoxicity seen with other vectors (Lukashev et al., 2016...along with a description of their experimental protocol. The three plasmids which have been deposited ...Gonçalves and Ventura labs’ plasmids, including protocols, check out the plasmids at Addgene: pAdSh.PGK....

...published a protocol that would work for your system. Addgene has depositor submitted protocols and links... Check out our CRISPR Guide Pages View CRISPR Protocols      ...

...of the genome and located immediately 5’ to a protospacer adjacent motif (PAM) sequence. Co-delivery of...a given phenotype. Image Credit: www.flickr.com/photos/72005145@N00/5600978712/ What CRISPR-based ...Using Pooled Lentiviral CRISPR Libraries. Current Protocols in Molecular Biology 115: . https://doi.org/10.1002...