...the liquid. Molecules absorb different wavelengths of light to different degrees and many molecules have...spectrophotometer measures these absorbances using UV-transparent cuvettes. First you measure the absorbance of...the ladder. You then run samples of your DNA at different dilutions and quantify based on band intensities...elute and store your plasmid DNA Learn about the different grades of plasmid DNA preps Resources on Addgene.org...Addgene.org Visit the Molecular Biology Reference Find plasmids for your research Browse protocols for ...

...either from human or plant sources) in both the strength as a promoter and the activity to release gRNAs...heart disease, and type 2 diabetes, across four different types of human cells, including T cells and hematopoietic... interest with only minimal genetic payloads. Different from existing multiplex array architectures, including...when repetitive tRNA sequences must be avoided, different tRNAs with optimized 5’ leader sequences can be... lab of Xue Sherry Gao at Rice University.    References and Resources More resources on the Addgene blog...Adding Precision and Flexibility to CRISPR Editing References Yuan Q, Gao X (2022) Multiplex base- and prime-editing...

... interest, and a polyA signal. Created with biorender.com. ITR structure  ITRs are symmetrical 145-nucleotide...resolution site (trs) are noted. Created with biorender.com. ITR modifications and types  Since ITRs are...expression can be altered by using ITR sequences from different AAV serotypes that vary in their RBE and T-shaped...multiple AAV serotypes (Wilmott et al., 2019), and different combinations of ITR and capsid serotypes vary ...restriction digest methods.    Resources and references More resources on the Addgene blog Viral Vectors...Addgene.org Addgene's AVV guide AAV Viral Preps References  Earley, L. F., Conatser, L. M., Lue, V. M., ...

...not exert force by dynamically rearranging into different polymerization states. Instead, R body monomers...natural context or 3) be modified to extend at different pH levels. Domesticating R bodies We have shown...that R bodies can function to break membranes of different organisms. And finally, we demonstrated that R... can be “tuned” with mutagenesis to extend at different pH values. Using a screen that takes advantage...region produced R-bodies that could extend at different pHs. Plasmids expressing expressing both wild ...School. Follow her on Twitter @jessicapolka.     References 1. Kanabrocki, J. A., Quackenbush, R. L. & Pond...

... labs (see table below) have different PAM requirements. These different PAM requirements allow researchers...discovered all the time and the requirements of these different systems can vary (for example, xCas9 is a variant...neither bind nor cut the target. Cas9 homologs from different organisms or Cas9 mutants developed in a variety...researchers to target many different genomic loci. 3. Cleave target DNA resulting in a double-strand break...sequence.   Now that you know about all the different components of CRISPR, It's time to start thinking...

... is it different from a CV? Get familiar with the general format of a resume and the different presentation...If the job posting says “French-speaking is an asset,” mention “speaks French.” Employers usually don'...you volunteer to do science outreach and teach children about science? Or perhaps you have presented in... science outreach and writing was how I got my current job as a Communications Coordinator. Wondering ...

... Plasmid Reference Molecular Biology Reference Promoters Promoters Definition A promoter is...Addgene's molecular biology reference material covering promoters, transcription and the transcription...transcription of DNA into mRNA. Promoter binding is very different in bacteria compared to eukaryotes. In bacteria...promoter region. Each sigma factor recognizes different core promoter sequences. Operons Although bacterial...regulation, like operons. Operons are a cluster of different genes that are controlled by a single promoter... types of RNA polymerases that all transcribe different genes. RNA polymerase I transcribes genes encoding...

...approach. One central bottleneck is that most recurrently mutated cancer genes are rare and most of the...found in genome sequencing studies (as well as reference cDNA clones mapping to well-annotated cellular...signaling pathways as part of our Genetic Pathway Reference Set. These Gateway-compatible entry clones are...cancer mutant collection and the genetic pathway reference collections are described in this paper Systematic...Central PMCID: PMC5386591.  The genetic pathway reference collection is described in this publication ...Comprehensive Set of MAPK1/ERK2 Missense Mutants. Brenan L et al. Cell Reports, 2016. PubMed PMID: 27760319...

...help define the different cells present in a mixed population, or identify differentiation and disease states...essential for synaptic transmission. Cleavage of VAMP2 renders it inactive, thus inhibiting neurotransmitter release...some genes can change depending on the location. Current methods for visualizing the location and function...challenges such as the inhibition of immune cell differentiation. Furthermore, TALEN and ZFN require custom ... any immune lineage without affecting cell differentiation in a high-throughput manner. They designed ...

...These cells are then transfected with several different constructs for the reporter to work and to make... visual read out allows for easy screening of different gRNAs by simply changing the gRNA used when transfecting...labor-intensive, require multiple fluorescent proteins, and aren’t that efficient. To overcome these issues the Denham...concept the Denham lab compared editing at three different genomic sites in the AAVS1 locus using GEmCherry2...GEmCherry2. In this experiment they saw a clear difference in mCherry expression using FACs sorting, confirming...mutant of interest, give these methods a try.  References Højland Knudsen C, Ásgrímsdóttir ES, Rahimi...

...the answer isn't to work harder - it's to work differently.  Tools to Try  Checklists Found in surgical ...and your workflow can have a huge impact. Test different approaches, like whether it's better for you to...as a Do Not Disturb sign, even if the headphones aren’t actually playing music. Nontraditional cues are... more than anything else I tried, made a huge difference in my ability to focus, especially on days when...what can be a rather vicious cycle.  Everybody’s Different   The truth is, there is no One True Way to Error-Free...the detail-driven daily grind of science.      References and Resources Images used Roberson Lab, UPenn...

...help us in different ways and in diverse areas. It is helpful to have mentors from your current organization...that you are an expert networker (or if you just aren’t doing it at all yet) watch my webinar, “Not Networking...you identify specific skills you need for your current position Understand your interpersonal challenges...someone you trust. A good coach can make a big difference, if you can find one. Create your own peer mentoring...with progress. Tell them how their advice made a difference, helped you make a change for the good or resulted...

...targeting the donor and acceptor fluorophores to the different biomolecules you’d like to test for interaction...that the targeting specificity requirements are different for the donor and acceptor fluorophores. The donor...acceptors are delivered to the wrong address in a different city, you can still develop a successful FRET-...signal is unaffected  Due to these fundamental differences in targeting specificity for donor and acceptor...blogger James D. Fessenden! James D. Fessenden is currently an Assistant Professor in the Anesthesia Research...channels involved in skeletal muscle contraction.   References 1. Degorce, François, et al. "HTRF: a technology... theoretical aspects and recent applications." Current Chemical Genomics and Translational Medicine 3.1...

...ortho-AP1867) via various chemical linkers. Our current lead dTAG molecule is dTAG-13 due its immediate...selectivity, broad utility for degrading targets in different cellular compartments, and in vivo utility. dTAG...dTAG molecule toxicity in new cell lines Treat parental cells (or cells expressing a control -- for example...expression of these cereblon neo-substrates varies in different contexts (for example, IZKF1 and IZKF3 are cell-type...technology. Follow him on twitter @behnamnabet.   References Nabet et al. The dTAG system for immediate and... 14(5): 431-441. PubMed PMID: 29581585. This reference contains the full description of the dTAG system...

...progression, enzymes and metabolism reprogramming aren’t the whole story. It’s essential for a tumor to ... integrins (cell-adhesion proteins) targeting different organs. This means that exosomes not only promote...microenvironment’s malignancy but also incite different organs to become malignant. Exosomes also contain...guest blogger, Bárbara Pinho! Bárbara Pinho is currently a science communicator at the Portuguese science...issues. Follow her on Twitter @BrbaraPinho6.      References Allinen, Minna, et al. "Molecular characterization...stromal caveolin-1 expression predicts early tumor recurrence and poor clinical outcome in human breast cancers...

... for your specific scenario.    References and resources References Bates A, Power CA (2019) David vs...Constant domain. H = Heavy chain. Created with biorender.com     The modularity of antibodies means ...is relatively easy to physically divide up the different regions, both through genetic and proteolytic ...broken up, there are a few commonly available different fragments that retain antigen binding (Figure ...common antigen binding fragments. Created with biorender.com.   Why Fabs?  Antibody fragments have a ...

...good base from which to dive deeper!References and resources References Bendall SC, Nolan GP, Roederer M,...love FPbase’s Spectra Viewer tool for comparing different fluorophores. We also touched on fluorophores ...chemical coupling can be achieved through several different mechanisms. Most take advantage of chemical groups...interactions with other peptides. Created with BioRender.com.     Many biologists can lead perfectly ...PK (2012) A deep profiler’s guide to cytometry. Trends Immunol 33:323–332. https://doi.org/10.1016/j.it...

... disease, and much more! Resources and References References Wang, I. X., Grunseich, C., Fox, J., et ...replication process within the host. dsRNA viruses are inherently double stranded, while ssRNA viruses can generate... dsDNA viruses yield dsRNA through a slightly different mechanism thought to be associated with bidirectional...transcription, RNA is synthesized from a DNA template, inherently generating RNA:DNA intermediates in the process...blog CRISPR 101: RNA Editing with Cas13  RNA Interference in Plant Biology  ...

... put your experiments Nsync.   References and resources   References Dolan, E., Wang, Hou, Zhonggang,...while also highlighting their similarities and differences to Cas9. DNA targeting Cas enzymes Cas9 – The...Originally classified as Cpf1, Cas12’s primary difference from Cas9 is that it generates staggered breaks...(RNA knockdown, RNA editing, etc.). The main differences between Cas7-11 and Cas13 are 1). non-specific... Cas While Cas9 may be the most adaptable Cas currently available, the other Cas proteins should not be...

...override genetically encoded patterning. Based upon trends in studies over the past several years, it seems...negative) the action potential of a cell, the more differentiated and quiescent it is, and the more depolarized... toggled between open and closed states using different wavelength light pulses. Other resources from...bioelectricity, bioelectric researchers in many different fields came together to found the journal Bioelectricity...regeneration, medicine, and synthetic bioengineering. References Chernet, Brook T., et al. "Use of genetically...for regenerative and synthetic bioengineering." Current opinion in biotechnology 52 (2018): 134-144. PubMed...