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... 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...

... 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 ...

...tool is the endoprotease Botulinum Neurotoxin B (BoNT/B), which cleaves the synaptic protein VAMP2, a ... split the proteolytically active light chain of BoNT/B into two fragments (BotNT-N and BotNT-C) and joined... into pXPR_053. They expressed this construct in bone marrow stem cells from Cas9-expressing mice, and...

... 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...

...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...

... 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...

...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. ...

...vector backbone from the Shih lab. You can find more lentiviral expression vectors (empty backbones) here...Plasmid #52961) This is a 3rd generation lentiviral backbone from the Zhang lab expressing S. pyogenes Cas9...

...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...

... 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 ...

... 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 ...

...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...

...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...

...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...

...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 ...

...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...

...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...

...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.   ...

...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 ...

...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 ...