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We narrowed to 312 results for: bon;

Showing: 31 - 60 of 312 results

  1. Pooled Library Amplifications

    Type
    Blog Post
    ...lentiviral backbone might be more prone to recombination, while a library with a large backbone might be...consider if there are any features of the library backbone that would make the library more difficult to ...other colonies containing smaller, recombined backbones that may replicate faster. For this reason, if...

  2. MXS Chaining

    Type
    Blog Post
    ...Kaiser AD, Kornberg A. Enzymatic synthesis of deoxyribonucleic acid. VIII. Frequencies of nearest neighbor...neighbor base sequences in deoxyribonucleic acid. The Journal of biological chemistry. 1961. March;236:864–...Trautner TA, Kornberg A. Enzymatic synthesis of deoxyribonucleic acid. XI. Further studies on nearest neighbor...neighbor base sequences in deoxyribonucleic acids. The Journal of biological chemistry. 1962. June;237:1961...

  3. Plasmids 101: Control Plasmids

    Type
    Blog Post
    ...Plasmid A (encoding shRNA against human Gene X in backbone Y) was transfected into human cells. From this...control vector would be Plasmid A sans shRNA, or backbone Y alone. The empty vector control allows you to... vector control shows any effect of the vector/backbone itself on gene expression in your target cells... this experiment could be an shRNA—in the same backbone as Plasmid A—that does not target any mammalian...

  4. Plasmids 101: Biotinylation

    Type
    Blog Post
    ...and is the process by which biotin is covalently bonded to a specific lysine at the active site of a newly...extremely strong affinity for one another. Actually the bond between them is one of the strongest noncovalent...protein-ligand interactions found in nature. Once the bond is formed it is unaffected by changes in pH, temperature...PMCID: PMC4884002. 10. Boer, E. de, Rodriguez, P., Bonte, E., Krijgsveld, J., Katsantoni, E., Heck, A., …...

  5. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

    Type
    Blog Post
    ...respective species. Addgene carries >30 “empty gRNA” backbones which contain a plant pol III promoter and gRNA...plants and many of the aforementioned empty gRNA backbones also contain Cas9, which enables expression of...CRISPR-Cas9 components into plants. Nanoparticles (carbon nanotubes, Kwak et al., 2019), DNA nanostructures...in planta using chitosan-complexed single-walled carbon nanotube carriers. Nat Nanotechnol 14:447–455 ....

  6. 28 Hot Plasmid Technologies from 2015

    Type
    Blog Post
    ... resistances, restriction enzymes and plasmid backbones. pUCXKT contains a nonfunctional truncated version...site in pUCXKT, removing the stop codons during backbone digestion and providing the missing codons for... both kanamycin and ampicillin (from the pUC19 backbone). *False positives can occur in this system if...lentivirus production Finding a lentiviral vector backbone with just the right combination of promoter, cloning...Addgene, so on your marks...get set...RUSH. Boncompain et al., Nat Methods 2012. Mar 11;9(5):493-8. ...create the pCell-Free vectors, Gateway-compatible backbones which enable cell-free expression of proteins ...originating from cryptic promoters in the plasmid backbone (Giannaks et al 2003, Bert et al 2000). In 2000...

  7. A Needle in a Base-Stack: Cas9 Structural Biology

    Type
    Blog Post
    ... why: two arginines make base-specific hydrogen bonds to the guanines, but the N (a thymine in this case...case) is only bound by the phosphate backbone. These PAM bases must be present for Cas9 to unwind the ...from the CTD (yellow) make base-specific hydrogen bonds to guanines of the PAM in the non-target DNA strand... both lobes bind to the DNA and RNA phosphate backbones, allowing Cas9 to verify the structure of the ...

  8. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    ...cloning my target spacers into the PX330 vector backbone? A2: There is no need to add the NGG PAM sequence...designing oligos for cloning my target sequence into a backbone that uses the human U6 promoter to drive expression...plasmid having a targeting spacer cloned into the backbone (pX330 or PX335, depending if you want to use ...quickly check if the cloning of my oligo into a backbone like PX330 worked? A12: You can quickly check ...

  9. What's New in CRISPR - December 2019

    Type
    Blog Post
    ...tagging Lentiviral capsid-based Cas9/gRNA ribonucleoprotein delivery Cytosine deaminase for RNA editing...bioRxiv Find the ORANGE toolkit   Cas9/gRNA ribonucleoprotein delivery by lentiviral capsid-based bionanoparticle...bionanoparticle system for packaging Cas9/gRNA ribonucleoproteins for transient expression. They replaced the...

  10. An Addgene Summer Internship

    Type
    Blog Post
    ... cloning the Syntaxin3 gene in a pLenti vector backbone. During those five weeks, I worked with my mentors...had successfully incorporated Syntaxin3 into its backbone. Once we had gotten our results back from the ...successful clone of the Syntaxin3 in the pLenti backbone! Once everything was confirmed, I worked on my...

  11. New CRISPR Web Resources and #12DaysofCRISPR Recap

    Type
    Blog Post
    ... Cas13a degrades only the specified RNA target. Bonus: It displays similar on-target activity to siRNA...explains why pC016 - LwCas13a guide expression backbone with U6 promoter has already earned a blue flame...101: Epigenetics and Editing the Epigenome (plus bonus podcast with Addgene Advisory Board member Alex ...

  13. RNA Extraction Without A Kit

    Type
    Blog Post
    ... of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18:5294–5299...effect of ammonium sulfate on the activity of ribonuclease A. Biochimica et Biophysica Acta (BBA) - Enzymology...

  14. Antibodies 101: Fab Fragments

    Type
    Blog Post
    ...are covalently linked to one another by disulfide bonds between the two heavy chains and between one heavy... digestion that retains an antibody’s disulfide bonds within the hinge region. The resulting product is...fragments, which reduces the hinge region disulfide bonds, resulting in two independent Fab fragments with...

  15. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

    Type
    Blog Post
    ...MLV)-like particles loaded with Cas9-gRNA ribonucleoproteins (RNPs). According to the authors, they named...but the researchers was able to edit hiPSCs, mouse bone marrow (BM) cells, primary human hepatocytes, and...viral-derived Nanoblades loaded with Cas9-sgRNA ribonucleoproteins." Nature communications 10.1 (2019): 45. ...

  16. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

    Type
    Blog Post
    ... NGS result to a reference sequence to confirm backbone elements.   Confirming the gene/insert by aligning...NGS result to a reference sequence like a known backbone sequence. We’re not necessarily expecting a perfect...mismatches in the origin of replication or other common backbone elements. Since we’ve successfully grown the plasmid...

  18. Educational Planarian Tools for Teaching Developmental Biology Techniques

    Type
    Blog Post
    ...species. These plasmids all have a PR-T4 vector backbone, which is a modified version of L440. This vector...cloning to insert the cDNA into the desired plasmid backbone. The Sánchez lab plasmids allow students and new... gene of interest into the empty PR-T4P vector backbone. The empty vector PLL4, which is similar to PR-T4P...

  19. Plasmids 101: 5 factors to help you choose the right cloning method

    Type
    Blog Post
    ... of the essence. This method requires a vector backbone containing a topoisomerase I cut site, toposisomerase...the topoisomerase I generated overhangs in the backbone. Once combined, the cut and ligation processes...out protocols for plasmid cloning Find vector backbones Read our molecular biology reference ...

  20. QC Sequencing Technologies at Addgene

    Type
    Blog Post
    ... bases (A, C, T, or G) is supplied as a dideoxyribonucleoside triphosphate (ddNTP) so that after its addition...not leave much flexibility for discrepancies in backbone sequences. Most theoretical sequences submitted...deposits that contain many plasmids within the same backbone. The flexible auto-alignment, plus additional ...

  21. Avoiding the Dark Side of Fluorescent Protein Fusions with mOX FPs

    Type
    Blog Post
    ...modifications including glycosylation, disulfide bond formation, and proteolytic cleavage. FPs localized... cysteine residues form inappropriate disulfide bonds in the secretory pathway (Figure 1) [4,6–8]. Additionally...formation of inappropriate inter-chain disulfide bonds between FPs blocks Golgi complex-localized fusions...

  22. Plasmids 101: E. coli Strains for Protein Expression

    Type
    Blog Post
    ...track: Compatibility: Double-check your plasmid backbone and expression strain to make sure they are compatible...contain hydrophobic regions or multiple disulfide bonds may aggregate and become insoluble. These insoluble...below or browse Addgene’s collection of empty backbones.   Our Most Popular Plasmids 101 Posts: Origin...

  23. Using Phosphoserine to Study Protein Phosphorylation

    Type
    Blog Post
    ...phosphomimetic mutation. This emphasizes the advantages of bona fide pSer-incorporation into recombinant proteins...interest (Figure 2). 1. The phosphoserine OTS The backbone of the phosphoserine OTS, Sep-OTS, contains the...using the PhosTag system (see below). This plasmid backbone can also be cloned with an ORF of interest for...

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

    Type
    Blog Post
    ...CRISPR machinery, Quadros et al. recommend ribonucleoprotein (RNP) complexes of Cas9 protein and the separated...alleles using long ssDNA donors and CRISPR ribonucleoproteins.” Genome Biol. 18 (2017) (1):92. DOI: 10.1186...Homology Directed Repair Learn about Cas9 Ribonucleoprotein (RNP) Delivery Resources on Addgene.org ...

  25. Deep Dive: Statistical Tests (Comparisons)

    Type
    Blog Post
    ...to reduce false positives within a test.   The Bonferroni Correction and the Benjamini-Hochberg procedure...control for large numbers of comparisons. The Bonferroni does this by establishing a significance threshold...However, if many comparisons need to be performed, Bonferroni and Benjamini will greatly reduce family-wise...

  26. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    ...expressed from different plasmid backbones. They noted that backbone differences such as the order, or...used to assess how different lentiviral plasmid backbone elements impact viral titer and to compare different...

  27. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    ...chemically modify target bases without breaking the DNA backbone, making them efficient and precise tools for altering...from the target base and replaced it with a double-bonded oxygen. This simple enzymatic reaction enabled ...which incorrect nucleobases are removed from the backbone, creating abasic sites for further repair.   ...

  28. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    ...chemically modify DNA bases without breaking the DNA backbone, sidestepping the problem of HDR vs. NHEJ entirely...they purified HF-BE3 protein for delivery in ribonucleoprotein particles (RNPs) to both zebrafish embryos...editing Rees et al., 2017 HF-BE3   HF-BE3 and ribonucleoprotein delivery decrease BE3 off-target activity ...

  29. 15 Hot Plasmids from 2017

    Type
    Blog Post
    ...they purified HF-BE3 protein for delivery in ribonucleoprotein particles (RNPs) to both zebrafish embryos...components are provided in Golden Gate compatible backbones that can be combined with your gRNA and repair...been identified. In Rauch, et al. 2016 the Joseph Bondy-Denomy lab at UCSF identifes four proteins from ...
Showing: 31 - 60 of 312 results