...the wrong everything else.      References and Resources References Boston University Flow Core. Fluorochromes...standard curve, another quick experiment, or using a different vial and saving the poor, abandoned antibody for...assays involve multiple antibodies, each with a different signaling molecule, that all need to be detected...   Fig 2: Try to inversely pair fluorophore strength with protein abundance.    Second, you’ll ...

...Linearized Plasmids After PCR). Current site directed mutagenesis methods Current SDM methods rely on in vitro...those to PCR amplify the template plasmid. The parental, nonmutated DNA is digested with methylated DNA-specific...common methods for site directed mutagenesis   References 1. Trehan, Ashutosh, et al. (2016). “REPLACR-... on Addgene.org Browse Our Molecular Biology Reference Pages Choose Your Perfect Plasmid Backbone Find...

...journal.pgen.1004086 Ghezraoui H, Piganeau M, Renouf B, Renaud J-B, Sallmyr A, Ruis B, Oh S, Tomkinson AE...repair are easily re-cleaved while indel products aren’t, so repeated cycles will favor accumulation of ...Comprehensive Cancer Center at UNC Chapel Hill.   References Bennardo N, Gunn A, Cheng A, Hasty P, Stark JM...2014) Is Non-Homologous End-Joining Really an Inherently Error-Prone Process? PLoS Genet 10:e1004086 ....

...important that you sequence the final product. Reference Page Top Index...

...protein expression journey! Resources and References References Nabet et al. The dTAG system for immediate...expression of essential proteins (when knockouts aren’t viable). siRNA or RNAi are small, double stranded...days. There’s also a CRISPR for that! CRISPR interference (CRISPRi) targets a catalytically dead Cas9 ..., and expression is tunable by titration with different amounts of tetracycline. Tetracycline regulation...

...dependence on excitation power is important since different fluorescence imaging strategies use widely varying..., TIRF or spinning disk confocal use entirely different excitation power regimes. Therefore, it is unpredictable...the best way to compare the photostability of different fluorescent proteins? Measuring photostability...excitation power. By repeating these measurements for different fluorescent proteins and by comparing how the ...measurements reported in the literature are performed in different ways. We describe a number of issues with some...

...1013 - 1014 vg/kg, a dosage comparable to those currently used in human gene therapy trials. They further...reproducibility as scientists can assemble parts from different labs and experimental systems in their own research...mutants, determined protein expression levels from different promoters, and inhibited gene expression using...C-terminus of Cre (CreC) and the N-terminus of Cre (CreN) are fused to the truncated Phytochrome B (PhyBΔ...eye, muscle and epithelial cells, as well as in different stages of development, ensuring the breadth and...

...reduced-in-size options often have reduced binding strength, half-lives, and/or specificity as well. They’...bispecific, meaning that each scFv can target a different antigen; binding of each can be either simultaneous...convert anyone to downsize their affinity reagents, trendy as that may sound. Validated antibodies are still... a scientist looking for that perfect fit!  References and Resources Additional resources on the Addgene...AddgeneBrowse our recombinant antibody plasmid collection References Grumezescu, A. M. (2018). Drug targeting and ...

...Emily P. Bentley in October 2024. References and Resources References Demanèche, S., Bertolla, F., Buret...functions of natural plasmids. Created with BioRender.com. Resistance (R) plasmids: microbial defense... also causes outbreaks of food poisoning, the difference is virulence plasmids! This category includes...Mclellan, S. L., Willis, A. D., Comstock, L. E., & Eren, A. M. (2024). A cryptic plasmid is among the most...

...mammalian plasmid here and edit on! References and Resources References Kwart, D., Paquet, D., Teo, S. et...sequence constraints, or at least has looser or different requirements. Luckily, there are some available...necessarily more flexible than NGG; they’re often just different (e.g. NGAG, NGAN, etc.). Additionally, alternative...al). Additionally, SpG Cas9 with the same PAM preference of NG was isolated by the Kleinstiver Lab (read...

...28032891. This is a different version of Split-BioID. BirA* is fragmented at a different site than in Schopp... This can artificially select against PPIs that aren't stable outside the native cell context. The Split-BioID...of the BirA* fusion matters! Schopp et al saw different levels of fusion protein expression and biotinylation...research? Plasmids are available from Addgene! References Schopp, I.M., Ramirez, C.C., Debeljak, J., Kreibich...

...whenever you hear it (whether at a scientific conference or a karaoke bar), you probably want to sigh....methyltransferase 1 (DNMT1) to copy methylation marks from parental to daughter strands of DNA. When dCas9 is bound...nearly 100% demethylation without the confounds inherent to dCas9-TET, allowing one to study the causal... CRISPR-based tools for epigenetic editing.   References and resources:  Additional Resources on the Addgene...Addgene.org Read our CRISPR Guide to Epigenetics References Charlton J, Jung EJ, Mattei AL et al. (2020) ...

... can only carry around 4.5 kb. Different AAV serotypes have preferences for specific cell types. For example...end of an experiment series would behave very differently (distracted) from the ones tested at the beginning...Experiment Behavioral scoring systems (experimenter differences in behavior observation); experimenter blinding...viral service for their behavioral studies. References Stoica L, Ahmed SS, Gao G, Sena-Esteves M. Gene...

...or undetectable FRET, well-characterized FRET reference standards can be used to validate FRET measurements...sophisticated questions about cellular processes. Currently, FRET experiments can probe protein-protein interactions...Borghi 2016) to a selected protein. Overcoming current limitations to permit the use of multiple FRET ... diseases and even assist in drug discovery. References 1. Baird, Geoffrey S., et al. "Circular permutation...biomolecular structure and dynamics in live cells." Current opinion in structural biology 34 (2015): 52-59....

... to address the unique needs of students from different departments across the university and alterations...track postdocs nationwide. On this level, FoR is currently compiling a database of institutional responses...implementation of new strategies designed to cope with current increases. These two rulings along with progress...Howell -Founder of Howell Communications Philip Brenner, PhD - Assistant Professor of Sociology. University...public policy. Follow David on Twitter @driglar.   References: 1. Alberts, Bruce, et al. "Rescuing US biomedical...

...Several protocols have been written outlining the different steps and materials required for each [1, 2]. ...discussed above, each opsin variant has slightly different activation/inactivation kinetics. For example,...Biology from the University of Michigan and he is currently a Postdoctoral Fellow in the lab of Mary Jeanne... or follow him on Twitter @derekpsimonphd.    References 1. Gutierrez DV, et al. Optogenetic control of...channelrhodopsin for activation of CNS neurons. Current protocols in neuroscience / editorial board, Jacqueline...

...Find the MagLOV plasmid here!   References and resources References Duret, G., Polali, S., Anderson,...just be an artifact and eliminate sources of interference, but the result persisted. (Even better, multiple...be developed from other fluorescent proteins.  Currently, the team is working to identify how MagLOV’s ... of many optogenetic tools. And, since MagLOV’s parent protein AsLOV2 has already been adapted into a ...

...However, if your flow cytometer reads them out on different lasers, you should not need to compensate between... Guide Chart.  Pro tip! Some antibodies go by different names depending on your flow cytometer’s setup...look at the other channels you’re using first. A different fluorophore channel may be affecting your populations...first before you apply compensation values to your current channels. A notable exception If you have a sample...CD8+ T cells and cancer immunotherapies. She is currently a research technician in Arlene Sharpe's lab at...

...previous Addgene CRISPR publications.  References and resources References Pyhtila, B., Kasowitz, S., Leeson...with CRISPR pooled libraries, although we used different categories (Figure 2C). Note that in Figure 2,...through the years.  Libraries are divided into different categories based on function: knockout (KO), activate...have CRISPR plasmids deposited from labs in ~40 different countries, and we’ve shipped CRISPR plasmids to...

...antibodies, context is key.    References and Resources References Evan GI, Lewis GK, Ramsay G, Bishop...cell proliferation and tumorigenesis.  Using two different synthetic peptides corresponding to linear sequences... or the C-terminus, Evan, et al. isolated six different monoclonal antibodies (Evan, et al. 1985). Three...-9E10 variable regions and sticking them onto different scaffolds. But over the years, several alternative...