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TypeBlog Post...in cells, and without something to block RNase activity upon cell lysis, RNA is degraded. To effectively...-based solutions that work by inhibiting RNase activity within cells or tissues- they don’t actually chemically...A (1974) The effect of ammonium sulfate on the activity of ribonuclease A. Biochimica et Biophysica Acta...
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Plasmids 101: CcdB - The Toxic Key to Efficient Cloning
TypeBlog Post...the insert. Gateway® technology (developed by InvitrogenTM) is essentially a more modern version of this...CcdB-resistant strain is ccdB Survival™ from InvitrogenTM. Using either DB3.1 or ccdB SurvivalTM, one ...a variety of useful resources and protocols! We invite feedback from all members of the research community... -
Working with Nuclear Receptors
TypeBlog Post...sensors for a diverse set of fat-soluble hormones, vitamins, and dietary lipids. Included in this family are...progesterone, androgens, and estrogens), fat-soluble vitamins A and D, thyroid hormone, fatty acids, oxysterols...transfection platform for assaying transcriptional activity, for example, was a key development in the functional... -
Engaging with science and society at pgEd
TypeBlog Post...personal identity. We do this through a variety of activities, including creating educational materials and...addition to events that we put on, we are often invited to present our materials or speak about our experiences...our science communication posts Extracurricular activities for a strong science career path Resources on... -
BeHeard Award 2018: Diseases of Glycosylation, Arginine Mutagenesis, & Neural Development
TypeBlog Post...characterized by deficient phosphomannomutase (PMM) activity due to mutations in the PMM2 gene. Videira’s group... Decades ago, other labs discovered that the in vitro functions of 11 different Arg-mutant enzymes could...phenotypes. Developing a rapid test for the functional activity of VUSs will enable more accurate diagnosis of... -
Split-BioID: An Improved Method for Studying Protein-Protein Interactions
TypeBlog Post...interact. When this interaction occurs, BirA* activity is restored, and nearby proteins are labeled with...BirA* enzyme. When this interaction occurs, BirA* activity is restored, and nearby proteins are labeled with...proteins Y and Z part ways, BirA* biotinylation activity ceases. After biotin tagging of proteins in both... -
Plasmid-based Recombinant Monoclonal Antibodies: What They Are and Why You Should Be Excited About Them
TypeBlog Post...are monoclonal antibodies that are generated in vitro using synthetic genes that are typically expressed...process of antibody selection and maturation in vitro by selecting for antigen binding from libraries ...or mammalian display (Tsuruta et al., 2017). In vitro selection allows scientists to create antibodies... -
Adapting Toehold Switches to Detect Zika Virus
TypeBlog Post...sequence (Green et al, 2014). The second was an in vitro paper-based platform that allowed gene circuits ...more stringent, and it avoided any days of lost productivity waiting for follow-up DNA oligos to arrive. ...parallel. More broadly, we drew on the combined creativity and expertise of the group and yet never hesitated... -
Cpf1 Update: Comparison to Cas9 and NgAgo
TypeBlog Post...-base sequence. Double mismatches ablated Cpf1 activity, except when they were present in the 3’ end of... as Kleinstiver et al. saw no decrease in Cpf1 activity with 4-6 base deletions at the 3’ end of the target...found that both orthologs show lower off-target activity than that previously observed with SpCas9. Kleinstiver... -
Common Injection Routes in Mice
TypeBlog Post...Intraperitoneal (IP) injections are given in the peritoneal cavity, or the space surrounding organs in the abdomen...organs. Restraint: scruffing Location: peritoneal cavity Gauge: 25–27 Volume: < 2–3 ml Skill required: intermediate...tail vein) Intraperitoneal Scruffing Peritoneal cavity 25–27 < 2–3 mL Intermediate Intramuscular Mechanical... -
Viral Vectors 101: Producing Your rAAV
TypeBlog Post...cell-associated (Vandenberghe et al., 2010). For certain in vitro and in vivo applications, you can consider using..., E., Hernandez, A., Boye, S. L., Boye, S. E., Levites, Y., … Golde, T. E. (2020). Utilizing minimally...assay for adeno-associated virus vectors with sensitivity sufficient to detect single infectious events... -
Fluorescent Proteins 101: Fluorescent Protein Timers
TypeBlog Post... (500 to 580 nm) over the course of 18 hours in vitro (see figure 1B in Terskikh et al.) and even displays...showed that their versatile timer could be used in vitro, in mammalian cells, in C. elegans, and in Xenopus...changes in oxygen, temperature, and pH, but timer activity can be calibrated to these different conditions... -
CRISPR 101: Mammalian Expression Systems and Delivery Methods
TypeBlog Post...Plasmids containing gRNA and Cas9 are used in in vitro transcription reactions to generate mature Cas9 ...ribonucleoprotein complexes Purified Cas9 protein and in vitro transcribed gRNA are combined to form a Cas9-gRNA...nor are these methods limited exclusively to in vitro cell culture. The user should review the current... -
Isolating Genomic Regions of Interest with the CRISPR System
TypeBlog Post...analyzed, the Fujii lab also developed another "in vitro" form. In this form, the CRISPR complex, consisting...biotinylated gRNA, were successfully used in this "in vitro" form. Any tag system can be used to isolate genomic... isolation of DNA fragments and chromatin by in vitro enCh IP technology using recombinant CRISPR ribonucleoproteins... -
Transferable Skills Guide: Identifying Your Transferable Skills
TypeBlog Post...teamwork, communication, writing, management, and creativity; I have developed these skills through school...not just “do the job”. When they ask about your creativity, your potential manager wants to hear a specific...resolution Organization Leadership Problem Solving Creativity Data analysis Listening Negotiation Career Planning... -
Better Dyeing Through Chemistry & Small Molecule Fluorophores
TypeBlog Post...to small-molecules: red-shifted spectra, ion sensitivity, photoactivation, etc. These impressive advances...of the genetically encoded tag, the speed and selectivity of the fluorophore attachment, the brightness...William Perkin. His discovery set off a flurry of activity and the majority of the classic small molecule... -
The Importance of a Fun Workplace: Company Culture at Addgene
TypeBlog Post... work responsibilities, we make team building activities a priority and have organized many events over...right in your breakroom. 5. Throw some physical activity into the mix It’s easy to sit around and just ...fast! Spread word of your plans by email, gong, evite, posters, megaphone, or your company’s intranet.... -
Overcoming the AAV Size Limitation for CRISPR Delivery
TypeBlog Post...editing has quickly become a popular system for in vitro and germline genome editing, but in vivo gene editing... (SaCas9, 1053 amino acids) displayed cleavage activity in mammalian cells. SaCas9 produced indels at ...SaCas9 did not display higher levels of off-target activity than SpCas9, confirming its suitability for in... -
CRISPR 101: Any Base Transversion Editors
TypeBlog Post...considerable A → G editing activity at most sites, in addition to A → C and A → T activity. To drive selective...introduce T → C and T → G edits, though with limited selectivity between those outcomes, with an average efficiency... -
Deciphering the Mysteries of Behavior with Viral Vectors
TypeBlog Post...networks involved in directly controlling muscle activity and coordinating movement, several additional ...develops an enhanced propensity for movement and activity called locomotor sensitization. This enhanced ... investigated this further and showed that the activity of a particular subset of excitatory neurons (...