...unlikely to be found in your gene of interest) and prevent incorporation of extensive sequences used for cloning...Translation of the standardized leader peptide prevents secondary structures in the mRNA encoding your...genes from a single transcript, or if you want to prevent read-thru transcription from resulting in the production...Adaptation of Enterococcus faecalis to daptomycin reveals an ordered progression to resistance." Antimicrobial...

... the HIV-infected reservoir, researchers have previously used histone deacetylase (HDAC) inhibitors to...the HIV-1 genome from every infected cell and 2. prevent reinfection of those cells. Kaminski et al. designed...HIV-1 cleavage allows T-cells to survive, but preventing re-infection would require sustained expression...infects over 250 million people worldwide. As we’ve previously seen in the CRISPR field, the lessons learned...

...sugar-containing molecules in molecular biology labs (for a review, see Simpson et al., 2015). Visit Addgene's MIcrobiology... have visible colonies growing on the membrane wherever an E. coli colony expressing a functional binding...  References 1. Nobrega, Franklin L., et al. "Revisiting phage therapy: new applications for old resources...Bases in Bacteria and Their Viruses." Chemical Reviews (2016). PubMed PMID: 27319741. 3. Simpson, David...

...modification was impossible. With the genome engineering revolution came epigenome-engineering tools - zinc finger...silent, they could identify the target genes of previously uncharacterized enhancers. The popular TALEN-...CRISPR activators and repressors are thought to be reversible once the effector is inactivated/removed from...KRAB induced silencing was transient and quickly reversed in culture. However, DNMT3A-induced methylation...

... activation and downstream signaling cascades (reviewed in Abreu & Levitz, 2020), including light-sensitive...the LOV domain blocks the enzyme binding domain preventing downstream signaling pathways. Light exposure...optimize the intensities of light sources used to prevent activation of both tools with each light source....  Finding the right information When using a previously published system in your own work, you’ll need...

...everything there is to know is a strange and exciting revelation. Science in any field can generally be applied...or not. Choose wisely This point ties into the previous one too. I would recommend applying for multiple...factors that can make or break a lab experience. Previous Addgene blogs have discussed the process that ... but I’ve learned the value in finding mentors wherever I go. This story by Dr. Erika Moore in Science...

...yeast, flies, and even plants! In this blog we will review common uses of MoClo and the kits Addgene offers...the MoClo YTK Kit specifically designed for S. cerevisiae is at your service (Lee et al., 2015). This kit...2017). In addition to additional elements for S. cerevisiae chromosomal integration and gene expression, ...Extensive and Flexible Engineering of Saccharomyces Cerevisiae.” ACS Synthetic Biology, vol. 12, no. 11, American...

... biomolecules each containing multiple binding crevices that can nonspecifically soak up your fluorophore...suitable for targeting FRET acceptors to proteins (reviewed in [Yan & Bruchez, 2015]). To get you started,...HTRF: a technology tailored for drug discovery–a review of theoretical aspects and recent applications....energy transfer as a spectroscopic ruler."Annual review of biochemistry 47.1 (1978): 819-846. PubMed PMID...

...of plasmid prep do you need? As we covered in a previous post on plasmid preps, not all plasmid preps are...purification kits make it easier than others to prevent quality issues. For example, some kits reduce the...weight of the package during shipping. Of the kits reviewed here, NEB’s Monarch®, QIAGEN’s QIAwave, and Milllipore... high level of consistency between lots, then reviewing their quality assurance documentation can help...

...case letter to the beginning of their insert abbreviation to specify what species it is. Example: ‘h’ ...amino acid mutation is to list the one letter abbreviation of the wild type amino acid immediately followed... Methionine (Met) followed by the one letter abbreviation of the mutated amino acid currently at that ...

...Annals of Internal Medicine published systematic reviews on an agent used to promote bone growth based on... open access as it does about the state of peer review. 8)     That same issue of Science also included...showing overwhelming support for open access to peer-reviewed journal publications, although many don’t necessarily...

...recent conversations with three lab heads have revealed that a combination of hard work, determination...How do you manage work-life balance? Tom Ellis reveals “I don't. Ask me again in five years and hopefully...Scientists Maintain a Good Work-Life Balance? Read the Previous Posts in this Series Advice for Choosing a Research...

... et al. "PP09. 1–2352: Systematic review of incidence and prevalence of spinal muscular atrophy (SMA)....S65. Additional Resources on the Addgene Blog Previous BeHeard Awardees Other Efforts to Support Rare...

...be completed and reviewed. The timeline therefore will be as follows:  Request review: 1–2 days Prep and...µL (5 mL).  Once submitted, the request will be reviewed by Addgene for legal approvals and technical compatibility...

...TAAGCAGAATTCATGTGGCATATCTCGAAGTAC-3'. For the Reverse Primer, the design is similar, but we need to use the reverse complement to get...primer and the downstream restriction site to the reverse primer. Next, we need to examine the DNA sequence...homology to the ORF). We now need to generate the reverse-complement of this sequence so that we can successfully...successfully amplify the ORF. You can generate the reverse-complement using existing software (a quick internet...others). If we put the sequence we chose for our reverse primer (5’-TGGCATATCTCGAAGTACTGAGCGGCCGCTAAGCA-...TGGCATATCTCGAAGTACTGAGCGGCCGCTAAGCA-3’) into this calculator we get a final Reverse Primer sequence of 5’-TGCTTAGCGGCCGCTCAGTACTTCGAGATATGCCA...digestion, you will need to use a phosphatase to prevent re-circularization of the vector. You should treat...

...analyses with the relevant figures from a literature review. No more hunting through a whole folder of papers...might include a confirmatory image to show that a previously used reagent performs well, or you might publish... one such research product which can convince reviewers of your expertise and insight. We recommend sharing...

...Utility. Annual Review of Pharmacology and Toxicology 55:399–417 . https://doi.org/10.1146/annurev-pharmtox-...Controlling Stimulation Some studies rely on delivering reversible or varied amounts of stimulation to receptors...

...sequence reverse complement tool: Enter a DNA sequence and the program will give you the reverse sequence...sequence, complement sequence, or reverse and complement sequence.  Filter DNA tool: Removes non-DNA characters...

... Need a quick review of the different immune responses? We got you! You can also review antibodies in ...Antibodies can bind to a pathogen and physically prevent it from entering a cell, a non-Fc-dependent effector...

... a single point mutation in the PAM sequence, preventing the Cas nucleases from re-identifying it (3)..... Addgene has empowered researchers to harness previous experimental successes and further develop the... FJM, Wolf YI, Yakunin JvdO, Koonin EV. Nature Reviews Microbiology. 2011 Jun; 9, 467-477. Bondy-Denomy...