...sequence of a thermostable DNA ligase-encoding gene." Gene 109.1 (1991): 1-11. PubMed PMID: 1756968. 7. ...nick-closing activity of bacteriophage T4 DNA ligase." Gene 76.2 (1989): 245-254. PubMed PMID: 2753355. 12. ...blood-borne bacterial pathogens by use of a novel 16S rRNA gene PCR-ligase detection reaction-capillary electrophoresis...restriction cloning), adaptor ligation for next generation sequencing (NGS) library construction, and circularization...fragments end-to-end without the exo/polymerase generation of annealed complementary regions. Why use a... does fidelity mean for a ligase? DNA ligases generally prefer fully Watson-Crick base-paired dsDNA substrates...as a correct base pair. DNA ligases have been generally found to have a higher discrimination at the upstream...

...horizontal gene transfer, which occurs when bacteria of different evolutionary lineages share genes that can...bacteria that occurs when bacterial cells regulate gene expression in response to changes in cell density... the ASM General Meeting (now Microbe) in 2013 in Denver, CO. While I had been to the General Meeting ... were very valuable when shared with the next generation of scientists. The connection between mentors... with experience mentoring in STEM, including Addgene’s Joanne Kamens. Expanding Binning Singletons to...Binning Singletons, Inc. Additional resources on the Addgene blog Browse our blog posts on scientific conferences...

...description of the VGS process and use cases in Human Gene Therapy. Existing methods for AAV quality control...Adeno-Associated Viral Genomes in Academic Settings. Human Gene Therapy Methods 30:195–205 . https://doi.org/10.1089...and tips Resources on Addgene.org Find AAVs at Addgene Learn more about Addgene's viral service Find protocols...we partnered with Seqwell to establish a next-generation sequencing (NGS) QC platform that allows us to...methods, we showed that the DNA extract is a heterogeneous mix of single-stranded and double-stranded species...interest.   While the VGS process isn’t new to Addgene, we are excited to share the procedures and Python...Vector Preparations by Single-stranded Virus Next-generation Sequencing. Molecular Therapy - Nucleic Acids...

... vector that is better suited for analyzing the gene’s function. The process is shown graphically in the...figure, in which we add EcoRI and NotI sites to Your Gene of Interest (YGOI) for ligation into a recipient..., you should use a DNA analysis tool, such as Addgene’s Sequence Analyzer, to allow you to identify which...restriction site to improve cutting efficiency. You can generally add any 6 bases, but you should ensure that the...with 18bp of homology to the ORF). We now need to generate the reverse-complement of this sequence so that...that we can successfully amplify the ORF. You can generate the reverse-complement using existing software...the final product. Additional Resources on the Addgene Blog Learn About Other Cloning Techniques Catch...

...amplification kit. In this case, regions of gene S and Orf1ab gene are amplified. RPA reactions also contain...amplification by primers that target the E and the N2 gene of SARS-CoV-2. If the Cas12a-sgRNA complex binds...like CRISPR. Figure 1: An overview of the general schematic of CRISPR/Cas based COVID-19 testing ...protocol for Cas13-based detection of SARS-CoV-2 genetic material (CREST) Sensitivity: 10 copies of RNA ...of temperatures varying between 10-50°C. Visit Addgene's COVID-19 Plasmids and Resources Many thanks ...Protocol for Cas13-Based Detection of SARS-CoV-2 Genetic Material. Cold Spring Harbor Laboratory. https:...20detection%20(updated).pdf Additional resources on the Addgene blog Read more about SHERLOCK and DETECTR Learn...

...also allows for screening scFv generated from the B cell antibody genes from diseased or vaccinated humans...   Some phage display libraries screen antibody genes from B cells of immunized animals, but it's possible....1791  Deal CE, Balazs AB (2015) Vectored antibody gene delivery for the prevention or treatment of HIV ...also allows for generations of scFvs against two antigens that are impossible to generate scFvs against ...probe, but is easier and cheaper to generate since it’s genetically encoded.   Translational applications... of a tagged protein. Resources on Addgene.org: Browse Addgene’s Antibody Plasmid Collection Find antibodies...-linker-VH and VH-linker-VL configurations can generate functional scFvs; however, some individual scFvs...

...Gibson assembly, imagine that you want to insert your gene of interest into a vector with a large tag at the...by using Gibson assembly, you can insert both the gene of interest and the tag sequences into the vector...CATCH enables one-step targeted cloning of large gene clusters. Nat Commun. 2015; Sep 1;6:8101. PubMed...the J. Craig Venter Institute in 2009. Here at Addgene we added this option to our drop down menu of common...which chews back the 5’ ends of the fragment, generating long overhangs which allows the single stranded...advantage is that this process makes it easy to generate wild type and mutant constructs at the same time...primer melting temperature greater than 48℃. Both Snapgene and NEB have tools that help you design primers...

...may be the hot new way to manipulate gene expression, but other gene manipulation systems remain valuable...important ways to spatially and temporally control gene expression, especially in in vivo models, and new...at least six generations. In addition to the neuronal pThy1-Brainbow constructs, Addgene also has two ...Brainbow plasmids available from Addgene! Find Brainbow Plasmids @Addgene: Brainbow 1.0, 1.1, 2.1 plasmids...coupling of Cre-lox with FLP-FRT recombination to generate additional recombination events, and the synthesis...and sea anemone mKate2.) They then successfully generated custom antibodies to each of these proteins and...analysis. To achieve even cell labeling, they generated farnesylated derivatives of the fluorescent proteins...

... provide a highly versatile means of delivering genes to many different cell types with minimal immunological...effects. AAVs are used in experiments targeting genes to specific cell types in the brain, mapping neuronal...Resources at Addgene.org Addgene's Viral Service Find Retrograde AAV Browse Optogenetics Plasmids ... to our viral vector pages on the Addgene website and on the Addgene blog to get some ideas for future...Boyden on Optogenetics, Neuroscience, and the Future of Neuroengineering Primer on Optogenetics Resources... Addgene recently attended the Society for Neuroscience 2017 conference in Washington DC (#SfN17). This...newbies might want to get into the field. Find Optogenetics Plasmids for Use in Neuroscience Research Neuroscience...

...were compared head-to-head for gene expression changes of non-target genes and RNA-seq of a nuclear (MALAT1...among the first batch of antibodies we produced at Addgene and, as you might suspect, it has been one of our... need a second GFP antibody?  Similarities:  Generated against full length GFP and converted to recombinant...always best to have options.      Fig. 2: Addgene’s western blot for mEGFP-HRas (from Plasmid 18662... [N86/8R] page for more details. Image credit: Addgene.   Find anti-GFP [N86/8R] here!   Andrews NP...