... with similar efficiency as compared to plasmid-based expression of Cas9/gRNA and can be used for most...experimental systems that may not be amenable to plasmid based methods. One of the most common techniques for delivery...gRNA RNPs to plants, as an alternative to plasmid based techniques, has been an area of intense study. Two...alleviate many issues that may arise when using plasmid based systems, most notably the ability to perform CRISPR... delivery of proteins enables efficient protein-based genome editing in vitro and in vivo." Nature biotechnology...

...different parameters that your gating strategy will be based upon. We’ll try to keep it short though, promise...SSC: scattering To digitally extract your cells based on their size, the cytometer provides you with two...lymphocyte population (10.7% of all recorded events) based on their expected size. The strong signals close...not of interest can still be gated out later. Based on personal preference, some people like to make...staining signals. For this, you’ll need to gate based on the proportion between H and A (or W and A) within...When you’re interested in measuring a population based on a fluorescent stain, you might be wondering which...

...has been started based on DETECTR technology with the mission to “provide a CRISPR-based platform on which...SHERLOCK and DETECTR demonstrate the power of CRISPR-based diagnostics. SHERLOCK- Specific High Sensitivity... DNA or RNA with atomolar sensitivity and single base pair mismatch specificity.  SHERLOCKv2 SHERLOCK ...isothermal amplification by RPA. When Cas12a-cRNA base pairs with the dsDNA of interest, the DNase activity...Wadford DA, Chen JS, Chiu CY (2020) CRISPR–Cas12-based detection of SARS-CoV-2. Nature Biotechnology. https...

...optogenetics, such as channelrhodopsins and tools based on light-oxygen-voltage (LOV) domain. They can also...interactions, we recommend using NIR split reporters based on miRFPs (Shcherbakova et al., 2016), since they...Filonov et al., 2013). FRET biosensors were developed based on miRFP760-miRFP720 FRET pair, including Rac1 biosensor...intensiometric calcium indicators NIR-GECOs were developed based on mIFP6. Learn more about the characteristics, ...Brightness in HeLa cells vs. iRFP713, % d Ref BphP-based FPs miRFP670 https://www.addgene.org/79987/ 642...720 98,000 6.1 97 Monomer 510 4.5 117 12 CBCR-based FPs miRFP670nano https://www.addgene.org/127443...

...system. Biosensors based on Nano-lantern technology are also very robust, with an eNL-based calcium sensor...RLuc8) with Venus. This luminescent technology is based on the principle of bioluminescence resonance energy...They also developed calcium, cAMP, and ATP sensors based on Nano-lantern, showing the technique’s versatility...

...time was not right for the development of phage-based therapeutics. After a brief stint at the venture...from developing therapeutics to developing phage-based tools for detecting pathogens in food products. ...biotechstart.org Read Sample 6's Paper on Bacteriophage-based Diagnostics Additional Resources on the Addgene...

... let the virions out. We recommend using a Tris-based lysis buffer and sonication.  The obtained virions...particles that can transduce cells, which can vary based on target cells and can be altered by freeze/thaw...experimental outcome and planning.  Both ddPCR- and qPCR-based titering assays measure physical titers, meaning...post.  We generally do not recommend using ELISA-based titering assays, since those will quantify both ...Inactivation of Adeno-Associated Viral Vectors by Oxidant-Based Disinfectants. Human Gene Therapy, 32(13–14), Article...

...gravity-based column plasmid maxiprep kit takes 1-1.5 hours to complete, while this syringe-based method...Compared to a commercial magnetic bead-based kit, this cellulose-based method was faster, cheaper, and had... Addgene follows a similar workflow as a column-based plasmid extraction kit. First, you lyse the bacteria...

...are many protein-based toxin-antitoxin systems. One of the most well known protein-based toxin-antitoxin...counter this, methyltransferases add methyl groups to bases at specific nucleotide sequences, preventing the...system that is regulated by antisense RNA, which base pairs to cognate mRNA to inhibit its translation...Crystal structure of the MazE/MazF complex: molecular bases of antidote-toxin recognition." Molecular cell 11.4...

...for studying colloidal gold (1).  The method was based on using a thin plane (sheet) of light generated...microscopes, optical sectioning of a specimen is based on discriminating the out-of-focus reflected light... combining Ultramicroscopy with tetrahydrofuran-based tissue clearing (13,14). A few years later, Karl...Deisseroth’s lab developed CLARITY, which is a hydrogel-based method that also allows antibody penetration for...recently developed AAV (adeno-associated virus)-based fluorescent labeling methods that can be used with...

...microbiome studies.  Find the MAGIC plasmids here Paper-based synthetic biology approach to analyze gut microbiota...microbiome, James Collins’s lab developed a paper-based method that can be used to detect microbes and biomarkers...As mRNA expression cannot be identified with DNA-based qPCR methods, the ability to detect mRNA transcripts...Takahashi, Melissa K., et al. "A low-cost paper-based synthetic biology platform for analyzing gut microbiota...

...slightly with either a 1-7 base poly(U)tail in tRNA-gRNA constructs or a 1-4 base tail from the second tRNA... cassettes (considering the gRNA itself is ~100 bases) limit the number of gRNAs that can be packaged ...Pseudomonas aeruginosa to cleave 4 gRNAs separated by 28 base Csy4 recognition sites from a polycistronic transcript...

...LoxP (locus of X(cross)-over in P1) sites are 34-base-pair long recognition sequences consisting of two...event. This event can have three general outcomes based on the location and orientation of the loxP sites...above, a genetic modification can be restricted based on experimental constraints. This has been used ...asymmetric spacer "NNNTANNN", where "N" indicates which bases may vary from the canonical sequence. Among these...

...options come in two categories: protein-based and chemical-based. Before selecting one, check your antibody...concentration. That makes the choice easy! Protein-based blockers include dry non-fat milk or serums like... of blocker, you can also use gelatin.  Protein-based blockers are inexpensive and easily made in the ... tip! If you get a speckled image, your protein-based blocker is either degraded or not completely mixed...mixed together.   Commercially available chemical-based blockers, while much more expensive, are consistent...to another. Since antibody binding varies wildly based on proteins, and their differing amounts of available...Smaller dynamic range; no multiplexing Protein-based blocker Degrades quickly Wet transfer Increased...

...years, deaminase-mediated base editing (cytosine base editor or adenine base editor) and reverse transcriptase-mediated...in Zhu et al., 2019). Both cytosine base editors and adenine base editors have been developed for plants...Belhaj et al., 2013). Nuclease dead Cas9 (dCas9) based activators (such as dCas9-VP64) or repressors (dCas9...

...Browse CRISPR Plasmids By Function Genome Editing Cut Base Edit Nick Prime Edit Transcriptional Regulation ...

... modifications (~1-50 base pairs). Small edits require as little as 30-50 bases for each homology arm,...arm, but keep in mind these numbers may vary based on your locus of interest and experimental system.  Synthesizing...  Synthesizing ssODNs longer than 200 bases can be difficult, thus dsDNA plasmids are generally preferred...Learn about Non-Homologous End Joining Learn about Base Editing Read 3 Tips to Improve HDR Efficiency ...

...process. Each variant is assigned a functional score based on its library frequency before selection compared...libraries of DNA fragments that contain variable bases (NNN) at any given codon site of interest can be... technique described in Wrenbeck et al is a PCR-based approach for generating a customizable comprehensive...primer set contains three consecutive randomized bases (NNN) at a given codon site and multiple primer ...Keith E J Tyo, and Timothy A. Whitehead. "Plasmid-based one-pot saturation mutagenesis." Nature Methods ...contain random synthesis errors, especially single basepair deletions. Therefore, Gibson assembly is better...codons that can be mutated? 88 codons, or 264 basepairs, was the maximum length tested by Wrenbeck et ...

...day, you should be happy to learn that many DNA-based reagents are named for soup ingredients! From pSOUP... intracellular RNA dynamics. Aptamers are 10-100 base nucleic acid oligos that bind with high affinity...that control the expression of coding RNA. Spinach-based riboswitches can be used as stable metabolite sensors...mimic of green fluorescent protein by fluorescence-based selection and directed evolution." Journal of the...PMC4244833. Kellenberger, Colleen A., et al. "RNA-based fluorescent biosensors for live cell imaging of ...

...scientific field-based solutions or open source solutions. In comparison to paper-based record keeping,...notebooks and researcher's needs The familiar paper-based lab notebook is still a standard approach to record-keeping... or brainstorm, but we have to admit that paper-based record keeping barely copes with the growing amounts...future? As most payable software solutions have user-based pricing, it is good to know how many of your lab...