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We narrowed to 772 results for: INA

Showing: 61 - 70 of 772 results

  1. Plasmids 101: Sequence and Ligation Independent Cloning (SLIC)

    Type
    Blog Post
    Published
    Sept. 26, 2024, 1:15 p.m.
    ...SLIC), eliminated many of LIC’s constraints by relying on the power of homologous recombination. SLIC ...reading Harnessing homologous recombination in vitro to generate recombinant DNA via SLIC. Li MZ, Elledge...only does this system not use site-specific recombination, it also doesn’t require a ligation step. Based...Based on the robust system of homologous recombination found in E. coli, SLIC is a cheap, standardized... incomplete plasmid looks like a homologous recombination intermediate to bacteria, so it can be transformed...optionally boosts efficiency with the homologous recombination protein RecA. (It’s effective with as little... branch); or by incomplete PCR, which skips the final extension step to retain products with single-stranded...

  2. Multiple Plasmids at a Low Price: Kits from Addgene

    Type
    Blog Post
    Published
    Sept. 14, 2016, 2:30 p.m.
    ...of extensive sequences used for cloning into the final translated products. Negative selection markers ...expression vectors additionally contain various combinations of protein tags (His, Avi, and GFP) as well ...blocker isolated from a designed repeat protein combinatorial library by in vivo functional screen." Scientific...production of a downstream peptide, the transcription terminators in this series are just the tool you need. The...this portion of the BIOFAB Plasmid Set contain terminators of varying strengths centered within GFP/RFP ...characterized in Cambray et al., 2013. These terminators can be used to precisely tune your polycistronic... for a researcher to quickly generate N- and C-terminal epitope tag fusions to a protein of interest. ...

  3. A Primer on Optogenetics: Introduction and Opsin Delivery

    Type
    Blog Post
    Published
    Sept. 10, 2015, 2:30 p.m.
    ... lambda sutures and subsequent determination of the proper coordinates of your brain region of interest...activation and the kinetics of opsin activity/inactivity. Dr. Deisseroth has also generously made many... as the vector core at the University of North Carolina) that offer his constructs. Importantly, control...open-reading frame (DIO). Upon coexpression with cre recombinase (either from a mouse line or viral co-injection...specificity dependent on the promoter of the cre recombinase while still taking advantage of the robust expression...identical for all of these surgeries. Identify coordinates through a rodent brain atlas or research papers...prior to using the virus. You may need to adjust coordinates based on your own method of identifying bregma...

  4. pSiM24: Simplifying Plant Genetic Engineering

    Type
    Blog Post
    Published
    Sept. 29, 2015, 2:30 p.m.
    ...Agrobacterium binary vectors Vectors designed for use in Agrobacterium are termed binary vectors because...depositor Indu Maiti has created a new and versatile binary Ti vector for both transient and stable gene expression...and E. coli. Like plasmids used in other systems, binary vectors are cloned and prepped in E. coli. However...co-cultivation with the target plant species. Thus, binary vectors contain two origins of replication, one...transferred to the plant, similar to viral vectors’ terminal repeats. It’s a lot of information to fit into...into a single plasmid, so most first-generation binary vectors are quite large (>10 kb.) Since transformation...scientists want to transfer large genes, smaller binary vectors with a larger cloning capacity would be...

  5. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

    Type
    Blog Post
    Published
    April 5, 2018, 12:42 p.m.
    ... 69090 pMJ915 MBP (N terminal); 6xHis (N terminal); 2xNLS (C terminal) 47327 pET-28b-Cas9-... 6xHis (C terminal); NLS (C terminal) 62933 pET-Cas9-NLS-6xHis 6xHis (C terminal); NLS (C terminal...62934 pET-NLS-Cas9-6xHis 6xHis (C terminal); NLS (N terminal)   Easi-CRISPR efficiency and delivery...marker, likely by breeding to a mouse line with a recombinase. If it turned out your chimeric mice did not ... a standard dsDNA repair template is used. The original design for creating a conditional allele, or floxed...that the T7 promoter will not be included in the final ssDNA template. Figure 2: Overview of ivTRT...stop codon!  Figure 3: How to design a C-terminal insertion with Easi-CRISPR. Potential guide sequences...

  6. Genetically-encoded Sparse Cell Labeling - A SPARC of Innovation

    Type
    Blog Post
    Published
    May 21, 2020, 1:15 p.m.
    ...the brain of Cre-expressing mice to trigger recombination and expression of a transgene (Lin et al., 2018...with precisely truncated attP sites to control recombination of the gene construct. A predictable number ...effector or retain the stop cassette following recombination. Image from Isaacman-Beck et al., 2019. Sparse... Sparse Predictive Activity through Recombinase Competition (SPARC), a genetic tool that allows for the... downstream gene expression. attP and attB, recombination sites. These sites are target sequences of PhiC31...where the stop cassette is removed during the recombination step, the effector is expressed. In cells where...expressed and the cells are not labeled. PhiC31, a recombinase. It specifically integrates an attP site into...

  7. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    Published
    July 31, 2020, 12:30 p.m.
    ...natural cytidine deaminases, they evolved the adenine deaminase ADAR2DD to deaminate cytidine and used... dRanCas13b (catalytically inactive Cas13) to target the cytosine deaminase. Three rounds of rational ...Originally published Nov 30, 2017 and updated Jul 31, 2020. Cas13 enzymes are quickly becoming major ...editor: a Cas13 enzyme fused to an RNA adenosine deaminase (ADAR). Such a system would convert adenine to...protein, they examined ADAR1 and ADAR2, which deaminate adenosine to inosine in RNA, creating a functional...functional A->G change. They fused ADAR deaminase domains (ADARDD) to dPspCas13b, but observed low RNA editing...the full ADAR2DD to further optimize cytidine deaminase activity. While RESCUE can perform C-to-U edits...

  8. Viral Vectors 101: Parts of the AAV Transfer Plasmid

    Type
    Blog Post
    Published
    Aug. 11, 2020, 1:15 p.m.
    ...of an AAV plasmid. Inverted terminal repeat (ITR) sequence Inverted terminal repeats (ITRs) are what makes...in them.  Lox sites Lox sites, in combination with the Cre recombinase, are used to spatially and temporally...loss, it’s best to propagate AAV constructs in recombination-deficient bacterial strains, such as Stbl3s....FLEX or DIO. Lox sites are recognized by the recombinase Cre. Cre first flips the sequence between the...excise the sequence between the second pair. The final result is a permanently flipped transgene. By restricting...the end of the AAV genome, just upstream of the final ITR.  polyA  The polyadenylation or polyA signal...the end of the AAV genome, just upstream of the final ITR and downstream of the WPRE. TL;DR? Watch our...

  9. CRISPR 101: Any Base Transversion Editors

    Type
    Blog Post
    Published
    April 1, 2025, 1:15 p.m.
    ...base editor (ACBE), they inserted the deaminase into the C-terminal domain of Cas9, which narrowed the editing...bystander edits. Their final tool, ACBE-Q (for an N108Q mutation in the deaminase), produced an average... (AAG). When adenine base editors (ABEs) were originally being developed for A → G edits, researchers ... MPG to an ABE would allow it to act on newly-deaminated inosines. Pro tip! The nucleoside inosine consists...further development. Figure 2: Mechanisms of deaminase-based adenine transversion editors. Red arrows...processes. Created with BioRender.com. The first deaminase-free base editor allows guanine transversions ...! All base editors up to this point relied on deaminase enzymes, either to directly accomplish the base...

  10. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

    Type
    Blog Post
    Published
    April 25, 2017, 2:30 p.m.
    ...wtGFP is illuminated with UV-violet light (~400 nm), the chromophore photoconverts predominantly to the ...activates and inactivates Dronpa is not ideal, since this means Dronpa is quickly inactivated and less photons...PA-GFP to display almost no fluorescence when illuminated with 488nm light. However, when UV-violet light... to a 100-fold increase in fluorescence when illuminated with blue light (1). Similar to PA-GFP, scientists...derived from DsRed and mRFP, respectively. When illuminated with violet light, both variants are photoconverted...but then further exposure to blue light also inactivates the chromophore, leading to a non-fluorescent...find a version of Dronpa that doesn't become inactivated as quickly. Padron is a monomeric, reversible...
Showing: 61 - 70 of 772 results