We narrowed to 593 results for: des.1

...has error rates that range from roughly 1 per 500bp to roughly 1 per 10 million bp depending on the polymerase...for Gibson assembly. The steps following primer design and the PCR process itself are very similar to ...the downstream steps. For this example, we will describe how to copy a cDNA from one vector into a new ...YGOI) for ligation into a recipient plasmid. Designing primers for PCR based cloning     The basic...: Do not cut within your insert Are in the desired location in your recipient plasmid (usually in ...examine the DNA sequence that we want to amplify and design primers that will bind to and replicate it. The...ends of the ORF and how these are used for primer design: Because we are cloning an ORF, we want to clone...

...centrifugation (lenti) Centrifugation Titration time ~1 day (AAV); ~1 week (lenti) 2–10 days Initial titers Higher...will help you decide which is right for you. Table 1: Mammalian versus insect cells for viral production... (about three to four days), this second round provides the added benefit of producing a stock P1 virus...

... fusions that retain their individual activities (1). Researchers also discovered the modular format of...systems could be combined effectively (3). In Figure 1 for instance, the DBD of the fusion protein shown ...ubiquitin system has been devised (8). References 1. Casadaban M.J., Martinez-Arias A., Shapira S.K., ...activation. Including a prey-only control also provides a baseline level of reporter activation. Vary ...

...and is recommended for use in western blots (Figure 1). If you prefer to produce your own antibody or want...are also available from Addgene!   Figure 1: Experimental validation for Anti-V5 [IPI-SV5-Pk1]...biophysical studies of protein degraders. Nat Commun., 15(1), 8885. https://doi.org/10.1038/s41467-024-52871-9...promising tool for future structure-guided drug design and studies of protein ubiquitination and degradation...CRBNmidi plasmid here!   Kroupova, A. et al. (2024). Design of a Cereblon construct for crystallographic and...specifically for C. elegans (Cao et al., 2024). The team designed coding sequences for mScarlet3 and mScarlet-I3...

...animal will repeatedly nose-poke at the active hole [1, 2].    Place preference/place aversion also models... can be explained primarily in two different ways 1) your hypothesis is wrong or 2) there is some problem...follow him on Twitter @derekpsimonphd.    References 1. Stuber, Garret D., et al. "Excitatory transmission...other words, your optogenetic experiment should be designed around a behavioral test in which your circuit...of mouse transgenics. Everything that has been described is intended for the mouse; however, all the steps....Diester, Ilka, et al. "An optogenetic toolbox designed for primates." Nature neuroscience 14.3 (2011)...

...the tag sequence flanked by homology arms (Figure 1). In the first step of optimization, we started off...form the ribonucleoprotein complex (csRNP, Figure 1). We use in-house purified Cas9 protein, which, in...standard immunocytochemistry (ICC) assays.  Figure 1: Three ingredients of epitope-tagging experiments....daydreaming about sunny days in the UK.   References 1. Bressan RB, Dewari PS, Kalantzaki M, Gangoso E, Matjusaitis...knock-in. Another good design tool can be found at Desktop Genetics .For ssODN design, remember to incorporate... systems, you will have to design your crRNAs using other web-based design tools (for C-terminus tagging... not work in pull-down assays: Epitope tagging provides a solution Proteins - the workhorses of the cell...

... this blog post. I separate sgRNA design tools into database and de novo solutions. Database tools allow...automated rational design. The first of only three such databases, EENdb (published 1 Jan 2013, no longer...community in order to succeed. With respect to de novo sgRNA design, I again separate it into two categories...follow him on Twitter @CMacPhD.          References  1. Ghorbal, Mehdi, et al. "Genome editing in the human...single-target design”, “multi-target design”, “off-target aware”, “high mismatch limit”, “approximate design”, “... “empirical design”, “single-PAM design”, and “multi-PAM design”. Trends: By the end of 2014, tools began...with the post-design process such as primer and plasmid design. Examples:  “multiplex design”, “multi-method...

...sequences are inserted into the genome. Figure 1: Self-excising cassette schematic from Dickinson et...):1028-34. doi: 10.1038/nmeth.2641. Epub 2013 Sep 1. PubMed. Find plasmids from this publication at ... the summer of 2013, a remarkable nine papers describing CRISPR/Cas9 genome engineering methods for C....America. To use this SEC-based system, first design and clone 500-700 bp homology arms for your gene...injected 60-80 animals per gene, they obtained the desired insertions on the first attempt for 6/7 genes. ...

... RNAs inhibit plasmid replication.   Figure 1: The replication problem. Iterons are repeated ...actually does, and neither plasmid is maintained (Figure 1).  The partitioning problem Plasmid incompatibility...Agrobacterium tumefaciens. Genes & Genetic Systems 77:1–9 . https://doi.org/10.1266/ggs.77.1 Pubmed PMID: ... Environmental Plasmids blog). Scientists have designed small, high copy incompatible plasmids that lead...Pubmed PMID: 25889267 Ebersbach G, Sherratt DJ, Gerdes K (2005) Partition-associated incompatibility caused...

...Check out Addgene's New Viral Service! References 1. Hay, R.J., Macy, M.L. & Chen, T.R. “Mycoplasma infection...contaminations.” Methods in Molecular Biology 946 (2013). 1-13. Pubmed PMID: 23179822. 5. Segura, M.M., Kamen,...Production, concentration and titration of pseudotyped HIV-1-based lentiviral vectors."Nature protocols 4.4 (2009...best to start from scratch. Obtain a vial of the desired cell line from a reputable distributor such as ...extreme changes in osmotic pressure and pH can destabilize the viral envelope. One drawback of the multiple...

...nLightR (Kagiampaki et al., 2023).     Figure 1: Properties of nLightG and nLightR. A, Structural ...Article 6. https://doi.org/10.1038/s41592-022-01488-1 More resources on the Addgene blog Fluorescent proteins...will need to be accounted for in your experimental design. Find nLight plasmids here! Next steps While nLightG...

...Rodríguez et al., 2011).  Genetics of SPARC Figure 1: SPARC and SPARC2 use the GAL4-UAS system along with... several common genetic tools in Drosophila (Fig. 1):  GAL4-UAS, a transcription factor binding site....References: del Valle Rodríguez A, Didiano D, Desplan C (2011) Power tools for gene expression and clonal...

...refreshed figures made using BioRender.com. Figure 1: PRIME editing Please check out the updated guide...update, we have added new sections to share and describe some incredible new CRISPR technologies developed...much-anticipated prime editing section and additions describing CRISPR’s ability to make large genome edits using...transposases or the Cascade-Cas3 system. We also describe newly engineered Cas9s with more flexible requirements...

...defined as (1) the struggle between two opposing forces or (2) the result of competing desires or (3) the...journey even when they know the ending.    Fig. 1: We all know that the superheroes win in the end. ...

...from distantly related organisms. In 2006, less than 1% of the genes deposited at Addgene were synthetic....synthetic biology is expanding rapidly.  References 1. Kunjapur, Aditya M., Philipp Pfingstag, and Neil ...of 173 known synthetic and natural genes.  In designing and optimizing their algorithm, they found that...

...and take a look at each of its parts. Figure 1: Parts of an AAV transfer plasmid. Parts shown in ..., ubiquitous expression, while the human synapsin 1 promoter is neuronal specific (Haery et al., 2019)...this post, you’ve got the knowledge to be able to design a construct for the successful and specific expression...

...following three strategies might help!     Figure 1: Species on species staining can be a problem because...same host species as your secondary antibody since 1) your secondary should not recognize Fab fragments...immunocytochemistry (ICC). Not all of the solutions described here may be as effective for other techniques,...

...(the Gal4/UAS system), and the third post will describe how you can make your own mutant flies. Find Drosophila...

...the journal-specific guidelines.  Peer-review step 1: The first read At the first read of the paper, I ...group of Prof. Mark Tester, where she focuses on (1) salt-induced changes in root-to-shoot ratio in Arabidopsis...of my review - "The manuscript by ZZZ et al., describes ... The authors share a wealth of data on ... ...Additionally, you can ask that data and resources described in the manuscript are accessible to the community...

...blotting, named for its inventor Edward M. Southern (1), can be used to detect DNA identity, size, and abundance...hours at low cost with restriction mapping (Figure 1). Though some labs may prefer to send DNA directly... serotonergic system and behavior.     References 1. Southern, Edwin Mellor. "Detection of specific sequences...the order of nucleotides in a string of DNA (7). Sanger sequencing uses modified nucleotides, called dideoxynucleotides...the widespread use of Sanger sequencing. Building upon this technology, radiolabelled nucleotides were...causes sickle cell anemia. The mutation happens to destroy a sequence recognized by the restriction enzyme...synthesis reaction along with all four regular nucleotides results in the synthesis of DNA strands of all...