...fragments are named according to the number of strands in the canonical beta-barrel structure. Most frequently...frequently, the proteins are split between strand 10 and 11 to yield large (1-10) and small (11) fragments...oligomerisation. Eastwood T, Baker K, Brooker H, Frank S, Mulvihill DP. FEBS Lett. 2017 Dan Mulvihill ...shifting tag (FAST) for use with green-yellow or orange-red fluorogen A split fluorescent reporter with...

...chain and light chain for the antibody and when translated into protein will assemble into a fully functional...synthesize the genes that encode those amino acids (Tran et al., 2016). Each antibody would then be tested...P, Frenzel A, Wong J, Koh XY, Eng H-Y, Lane D, Lefranc M-P, Clark M, Dübel S (2018) When monoclonal antibodies...:633–644 . https://doi.org/10.1083/jcb.201409074 Tran NH, Rahman MZ, He L, Xin L, Shan B, Li M (2016) ...

...fall within a 10 fold range. Plasmids that don't meet the lower bound of this range are “failed” and removed...mutations, we check to see if they affect the translated amino acid. We also confirm that the species ...2017) Genome-scale CRISPR-Cas9 knockout and transcriptional activation screening. Nat Protoc 12:828–863...

...Gate cloning, perform an exonuclease cleanup, transform, plate, and pick. In short, the Fragmid system...AAV pRDA_889 (ITRs from AAV2) Mammalian Transposon pRDA_757 (Piggybac; constitutive expression...   Drosophilia Plasmid pRDB_053 (for transgenic flies; gypsy insulator sequences; white + selectable... drosophila) Delivery type (lentivirus, AAV, transposon, Plasmid) Destination vector (dropdown options...

...sequences that, when bound by transcription factors, can increase a gene’s transcription. Sometimes enhancers ...and proteins that forms chromosomes. These long range DNA interactions are often detected with chromosome...control gRNA or predicted gRNA off-target sites. Long-range chromatin interactions identified with the multiplexed...

...would at least be guaranteed a successful result? While there are never any guarantees, some cloning techniques...Resources” section at the end of the post! Speed: Your grant deadline is approaching or your advisor is demanding...your insert, DpnI digest away the templates, and transform the PCR products into E. coli. Ligases within ...

...during ESC isolation. Low Chances of post-transplantation rejection when used therapeutically High Low...similar functional utility, iPSCs bypass the transplantation compatibility issues and ethical concerns of... used therapeutically, the chances of post-transplantation rejection are low. Further, as iPSC generation...somatic cell type to another, also known as transdifferentiation.   Many thanks to our guest blogger, Kusumika...

...NC: nucleocapsid RNA chaperone, RT: reverse transcriptase. Right: T1 tomato plants that inherited an edited...Cultured rat hippocampal neuron expressing pan-membrane ASAP5. Solid blue circle marks the initiation ...via the Kv2.1 targeting motif as well as a pan-membrane construct without the Kv motif. Altogether, if...kinetics and high responsivity near the resting membrane potential, ASAP5 is a great new option. Find ASAP5...

...production of dsRed E5. This timer predictably transitions from green to red fluorescence (500 to 580 nm... of vesicle fusion and release from the plasma membrane by fusing various vesicle cargoes to FP timer ...fusing Medium-FT to LAMP-2A, a lysosome-associated membrane protein, and following its subcellular localization...whether or not newly synthesized or long-lived transcription factors control gross changes in gene expression...

...blastn), Standard Protein BLAST (blastp), and Translated BLAST (blastx). NCBI has a terrific getting-started...gaps in the nucleotide sequence, you can run a Translated BLAST (blastx) search to check for expected ORFs..., due to the additional searches involved in translating the nucleotide sequence into all six possible...choose for your sequencing result and can easily translate it, we recommend using blastp over blastx. The...

...cases being reported (right, red), likely due to quarantining {% video_player "embed_player" overrideable...-examine our global data at a finer temporal granularity. For each paper, we observed that the demand ...imposed on scientists. Figure 6: Model fit. Orange line depicts observed data. Blue line is the sum...depending on when we believe the China COVID-19 date range is, they could be the most impacted or least impacted...

...expressing neurons and GFP (green) to label neurons transduced by each serotype. Working at Addgene has ...answers hundreds of AAV questions every year on a range of issues related to AAV vectors, many from people...S., & Klein, R. L. (2018). More expansive gene transfer to the rat CNS: AAV PHP.EB vector dose–response.../10.1038/s41434-018-0028-5 de Backer, M. W. A., Brans, M. A. D., Luijendijk, M. C., Garner, K. M., & Adan...

...visualize cell cycle dynamics, titrate inducible transgene expression, or a variety of other clever applications...expression) and a polyA signal (to terminate transcription) and the four constructs were then combined ...derived from the tyrosine-protein kinase Lyn) Membranes 3 mCherry 587nm/ 610 nm human β-Actin Actin ...their 6-mer sequences, are rarely found in the transcriptomes of these species and can often be used to clone...

...antibiotic resistance using multiple approaches. Major restaurant chains including Chipotle and Panera Bread have... joining machinery, so a CRISPR-induced double-stranded break (DSB) in the genome is lethal. If this DSB...diverse, and each phage has a narrow bacterial host range. To further potential clinical applications, Ando...Beisel, Chase L., Ahmed A. Gomaa, and Rodolphe Barrangou. “A CRISPR design for next-generation antimicrobials...

... elaborately pranking each other, dressing in costume, and holding competitions (ranging from cook-offs...orderly and timely fashion. 9. Ask for help My grandma used the old adage “many hands make light work”..., evite, posters, megaphone, or your company’s intranet. Don’t forget about the power of simple word of...

...particles that can transduce cells. Infectious titers are typically quantified by cell transduction assays. Wild-type....qPCR...Digital droplet PCR...DNA dot blot...Transduction assay...SDS-PAGE...Electron microscopy… Any ...titer of a viral vector stock can vary based on transduction target and can also be altered by freeze-thaw...

...that emerged from intensive engagement with a wide range of people at the University of Wisconsin in 2011...reproducibility an ELN should have integrations with a broad range of relevant generic and science-specific tools. ... however, in some cases it would be possible to grant those wanting to understand and/or attempt to reproduce...Massachusetts, and feels privileged to be part of the vibrant life sciences research and commercialization community...

... protein of interest, such as a sample transiently transfected or stably infected with the protein of ...downregulated, examine a protein’s stability or post-translational modifications, or study how a target protein...experiments using a tagged protein may not be translatable to the endogenous system and will need to be...

... package stays in transit for a certain number of days and tracking average transit time for each courier... often weren’t easily accessible, due to a wide range of logistical barriers. Addgene worked to identify...Protocols Agarose Gel Electrophoresis Bacterial Transformation How to Design a Primer Creating Bacterial Glycerol...

...popular papers, but looked for papers from a wide range of authors. Just start writing When I first started...drafts of the manuscript. This is the time to transform the document into something cohesive-- change ...Thompson, and Thomas D. Gilmore. "Histone acetyltransferases and histone deacetylases in B-and T-cell ...Tips for Improving Your Next Manuscript 6 Tips for Grant Writing Resources on Addgene.org Check out Our...