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...libraries containing combinations of different gRNAs in each plasmid. Each gRNA combination can be tracked ... an easy-to-use tool to analyze the complex combinatorial genetic networks underlying your favorite biological...screening are vastly useful for mapping the combinatorial genetics that underlie complex regulation in...barcoded gRNA libraries that can be used to combinatorially modify the genome, screen for a particular ...successfully applied to functionally characterize combinatorial gene knockouts generated via multiplexed gRNA...expression (Wong et al., 2016), in addition to the combinatorial expression of other genetic elements including... Figure 2: Strategy for assembling barcoded combinatorial gRNA libraries. (Adapted from Wong et al., 2016...

... or gene, as well as to study the effects of inactivating/activating mutations. In biomedical research.... In recombineering, bacteria express phage recombination machinery, which facilitates easy genetic modification...modification by homologous recombination. In most recombineering strategies, only 3-4 phage proteins are...a temperature-sensitive plasmid. Homologous recombination then joins the linear PCR products into stable...the recombineering plasmid is not maintained. Recombination efficiency is high enough to negate the need...own electrocompetent cells in which the phage recombination machinery is induced using L-arabinose. These... the steps of gel purification and in vitro recombination. Across all modifications described in Trehan...

...iGEM team originally used a guaB knockout E. coli strain to assay for functional decaffeination in cells...supplemented with various caffeinated beverages, but not with caffeine-free drinks. Finally, they demonstrated... pulse of blue light, and is reversible upon termination of light stimulation. CRY2clust enables faster... changes in intracellular Ca2+ levels than the original version containing CRY2 without the extra peptide..., termied soCoChR, by fusing a fragment of the kainate receptor subunit to the C-terminus of the protein...the group was able to demonstrate that while illumination of neighboring cells evokes action potentials...these APs are absent in soCoChR-expressing cells. Finally, using synaptic receptor blockers to distinguish...

...maintaining intensity at ~85% of its original value after 60 minutes of illumination, highlighting its reliability...scientists. Seeing inside these specimens has offered illumination on biochemical processes that are crucial in...interpretation of FRET-based experiments. A popular combination is Alexa594, and Alexa647 due to their brightness...

... and increase your lab sustainability. For my other posts on lab sustainability, check out my Recycling...This is the last post in a blog series on sustainable practices in the lab, where I’ll list some easy...unimportant. With so many big contributors to the sustainability problem, how can one little change make a difference...quick things above, try joining or starting a sustainability project. This could be: Setting up a recycling...Learn about other creative approaches for lab sustainability Resources on Addgene.org Sign up for our ...

...vectors, recombination often occurs between the long-terminal repeats (LTRs). After recombination, one resulting...intramolecular DNA recombination. This naturally occurring process is mediated by recombinases like RecA and...undergo recombination, two smaller plasmids are formed. Viral vectors are prone to recombination In lentiviral...replicate.   How can I prevent recombination in my samples? Plasmid recombination can affect viral vectors ...have been engineered to further reduce recombination. Recombinase-deficient strains like Stbl2, Stbl3, ... repeating elements. Knowing how to amplify recombination-prone plasmids and testing your prep can save..., and NEB stable E. coli reduce the rate of recombination in samples. For a more detailed comparison of...

... you encounter mycoplasma contamination. Causes of mycoplasma contamination The mycoplasmas enter the ...above. The contaminants can spread through dirty clothing, lab wears, human speech near the laminar airflow... risk of contamination. Mycoplasmas can spread from these sources through cross-contamination and due ... a source of mycoplasma contamination. Due to its turbid nature, contamination is difficult to be detected...during working in the laminar airflow are also among the leading causes of contamination. These aerosols generated...cell culture process and hence eliminate one of the causes of contamination. If possible, a separate lab...bacterial and fungal contamination. Proper PPE Since humans are the major source of contamination, the prevention...

...to avoid MMR: 1) E. coli with inactivated MMR: Using E. coli with inactive MMR is definitely the easier...minutes quickly inactivates the repressor and allows for expression of the recombination genes.  After ...confirmation of recombinant clones If a antibiotic resistance gene has been inserted, recombinants can first...genome engineering and is based on homologous recombination. It allows for direct modification of DNA within...called recombineering - short for homologous recombination-mediated genetic engineering.  It can be used... These enzymes then catalyze the homologous recombination of the substrate with the target DNA sequence...nucleotides of homology to the target site for recombination. Key components of the Lambda Red recombineering...

...plasmid-based recombinant monoclonal antibodies Resources on Addgene.org Find recombinant antibody plasmids...leading to problems with reproducibility and an inability to improve upon these critical research tools.... Addgene to create a much needed open-access recombinant antibody/affinity reagent resource for the neuroscience...and data, the platform can be expanded for any recombinant antibody tool and therefore will grow to serve... the entire scientific community.     Why recombinant antibodies? Animal-based methods for antibody...cryopreservation (Bradbury & Plückthun, 2015a, b). Recombinant antibodies are plasmid-based and offer numerous...monoclonal antibodies. You can learn more about recombinant antibodies in this blog post. Hello, NABOR! ...

...Fvasconcellos.     How are Recombinant Antibodies Made? To make recombinant antibodies, you first need..., 2015).    The Many Benefits of Recombinant Antibodies Recombinant antibodies offer numerous advantages...definition of recombinant antibodies are essential for good reproducibility. Non-recombinant antibodies ...predict a similar revolution for antibodies. Recombinant antibody technology makes it easy for scientists...components of the scientific toolbox. What are Recombinant Antibodies? Antibodies are used by scientists...scientific institutions have proposed a shift to recombinant DNA-based antibody technologies (Bradbury & Plückthun...single antibody recognizing one epitope. (Bottom) Recombinant monoclonal antibodies are encoded in plasmids...

...specificity in retinal cells. To do this, the group developed a new strategy for finding retinal cell-type ...monkey, and human retinas Next, the 113 AAV promoters that were active in mouse retinas were injected into...human retinas but not in mice was low (R=0.12), suggesting that mice could be used to eliminate promoters...gene expression in retinal cell types and classes in mice, primate, and human retinas. These synthetic ...types, such as inhibitory neurons, rods, cones, or retinal ganglion. Rather than using naturally occurring...they drove cell type specific expression in the retina.  Four approaches for designing synthetic promoters... with sequences upstream of the start codon of retinal cell-type specific genes from mice. Strategy #2...

...expression of other recombinases. Multiple orthogonal attB-attP pairs per recombinase may be arranged on...different recombinases are overlapping or nested on a register, then the operation of one recombinase on the... when treated with different combinations and orders of two recombinases. How might one read the state...they can control recombinase activity. For example, post-transcriptional recombinase activation via RNA...reduces the content of our paper, “Synthetic recombinase-based state machines in living cells” (1), into...explanation of how to adapt our engineered devices, recombinase-based state machines for your own experimental... about the order or timing of these factors. Recombinase-based state machines (RSMs), which take on a ...

...Having never been to China myself, I was pleased to be able to participate when the organizers of the...Dmitry Samarsky, and a very welcoming team from China’s Ribobio, the illustrious speaker list attracted...winners, one NIH Director and a co-founder of BGI-China presenting, it’s no wonder it was attended by over...were potential Addgene requestors and depositors. China is in one of the top 10 countries in number of plasmid...of Guangzhou and see some of the sites. Most fascinating to me was the outdoor market where one can shop...chemistry is complex and this work requires an interdisciplinary team.  Day two of the conference was kicked...from different species for genome engineering. The final keynote presentation was by Nobel Laureate Thomas...

...iGEM team originally used a guaB knockout E. coli strain to assay for functional decaffeination in cells...supplemented with various caffeinated beverages, but not with caffeine-free drinks. Finally, they demonstrated... pulse of blue light, and is reversible upon termination of light stimulation. CRY2clust enables faster... changes in intracellular Ca2+ levels than the original version containing CRY2 without the extra peptide..., termied soCoChR, by fusing a fragment of the kainate receptor subunit to the C-terminus of the protein...the group was able to demonstrate that while illumination of neighboring cells evokes action potentials...these APs are absent in soCoChR-expressing cells. Finally, using synaptic receptor blockers to distinguish...

...plasmid-based recombinant monoclonal antibodies Resources on Addgene.org Find recombinant antibody plasmids...leading to problems with reproducibility and an inability to improve upon these critical research tools.... Addgene to create a much needed open-access recombinant antibody/affinity reagent resource for the neuroscience...and data, the platform can be expanded for any recombinant antibody tool and therefore will grow to serve... the entire scientific community.     Why recombinant antibodies? Animal-based methods for antibody...cryopreservation (Bradbury & Plückthun, 2015a, b). Recombinant antibodies are plasmid-based and offer numerous...monoclonal antibodies. You can learn more about recombinant antibodies in this blog post. Hello, NABOR! ...

...Fvasconcellos.     How are Recombinant Antibodies Made? To make recombinant antibodies, you first need..., 2015).    The Many Benefits of Recombinant Antibodies Recombinant antibodies offer numerous advantages...definition of recombinant antibodies are essential for good reproducibility. Non-recombinant antibodies ...predict a similar revolution for antibodies. Recombinant antibody technology makes it easy for scientists...components of the scientific toolbox. What are Recombinant Antibodies? Antibodies are used by scientists...scientific institutions have proposed a shift to recombinant DNA-based antibody technologies (Bradbury & Plückthun...single antibody recognizing one epitope. (Bottom) Recombinant monoclonal antibodies are encoded in plasmids...

... multiple Addgenies and most recently updated by Ina Ersing in August 2023. Resources and references ...  Ina Ersing...absence of unexpected DNA contaminants. The data is also used to determine recombination rates in FLEX/DIO/fDIO...Endotoxin contamination in vector preparations can alter the immunogenic properties of the final product...mycoplasma contamination. To date, Addgene has never had a case of mycoplasma contamination. In the event...is performed on viral genomes isolated from the final AAV preparation. Sequencing results are analyzed...particularly in large animal studies. Endotoxin contamination is minimized by using an endotoxin-free plasmid...

... multiple Addgenies and most recently updated by Ina Ersing in August 2023. Resources and references ...  Ina Ersing...absence of unexpected DNA contaminants. The data is also used to determine recombination rates in FLEX/DIO/fDIO...Endotoxin contamination in vector preparations can alter the immunogenic properties of the final product...mycoplasma contamination. To date, Addgene has never had a case of mycoplasma contamination. In the event...is performed on viral genomes isolated from the final AAV preparation. Sequencing results are analyzed...particularly in large animal studies. Endotoxin contamination is minimized by using an endotoxin-free plasmid...

..., Diana, Dolly, Eddy, Emma, Eric, Gizela, Henry, Ina, Inna, Ishrat, Jane, Jason, Jessica, Joel, Jonathan... revenue to build reserves so the company is sustainable. The difference is what nonprofits do with their..., Lianna, Luke, Marcy, Mary, Max, Maya, Meghan, Melina, Meron, Michael Lo., Michael Le., Michel, Michelle...

..., Diana, Dolly, Eddy, Emma, Eric, Gizela, Henry, Ina, Inna, Ishrat, Jane, Jason, Jessica, Joel, Jonathan... revenue to build reserves so the company is sustainable. The difference is what nonprofits do with their..., Lianna, Luke, Marcy, Mary, Max, Maya, Meghan, Melina, Meron, Michael Lo., Michael Le., Michel, Michelle...