... of scientists. 9. Plasmids 101: Positive and Negative Selection for Plasmid Cloning by Jennifer Tsang... (ex: antibiotic selection and auxotrophy) and negative selection (ex: toxin-antitoxin systems and sacB...101: Transformation, Transduction, Bacterial Conjugation, and Transfection by Alyssa Cecchetelli The ...

... is no way a manager can be efficient without delegating and empowering the team to solve problems together...together (tactics for effective delegation will be covered in a future post in this series). Members ...at formal communications events or at informal gatherings, like at lunch. Many experts recommend using ...

...the inside of the neuron more positive or more negative.  Neuronal stimulation: Depolarization When the...Conversely, when the inside of the neuron becomes more negative it becomes less likely to fire and is said to ...hyperpolarized. The chloride pump NpHR brings in negative chloride ions which hyperpolarize the cell. It...

...we'll focus primarily on antibiotics against Gram negative bacteria. In future posts, we'll detail selection...ribosomal subunit; inhibits protein synthesis (elongation step) Bacteriostatic 10 ug/mL Chloramphenicol...inactivates or completely removes (in the case of Gateway cloning) one cassette. Counterselection allows ...

... positive), with these puncta turning red (GFP negative/RFP positive) upon fusion of an autophagosome ...RFP-tagged LC3∆G. Like the unmodified LC3, GFP-LC3 is conjugated to phosphatidylethanolamine (PE) and recruited...autophagy. The RFP-LC3∆G, however, can not be conjugated to PE due to a C-terminal glycine deletion, so...

...2016 Addgene’s research team has been actively investigating ways to improve viral vector production. Whether...Whether redesigning viral packaging plasmids, investigating different viral production strategies or developing...method to improve AAV yields. Although we only investigated 3 serotypes it’s possible that this approach...

...next generation the CRISPR construct needs to be segregated away to prevent continued production of new alleles... are identified by a phenotype caused by the segregating mutation. Selecting these plants is a learned...integrated into a CRISPR system, then tracking the segregation of the CRISPR construct could be achieved much...

.... These studies used SMRT sequencing, TA-based ligation and tagmentation-based methods for AAV genome ...works for self-complementary AAVs and the TA-based ligation and tagmentation preparation approaches described...AAV genome is single-stranded and both TA-based ligation and tagmentation require a double-stranded template...

...product and vector ligate; however, it can potentially rebind and nick the newly ligated DNA. Salt helps ...an extremely large insert. Since the standard ligation reaction is fairly quick, make sure you stay organized...

...NotI sites to Your Gene of Interest (YGOI) for ligation into a recipient plasmid. Designing primers for... In our example, we will use EcoRI and NotI to ligate our cDNA into the recipient plasmid. Remember to...final Reverse Primer sequence of 5’-TGCTTAGCGGCCGCTCAGTACTTCGAGATATGCCA-3’. Preparing the PCR product for...

... Facultative anaerobe Facultative anaerobe Obligate aerobe Number of chromosomes 16 3 4 ... is a non-pathogenic yeast that operates as an obligate aerobe. It’s also a methylotropic yeast, meaning...DOI: 10.3892/br.2017.941 Bernauer L., et al., Komagataella phaffi as Emerging Model Organisms in Fundamental...

...for PZA-resistant Mycobacteria tuberculosis. Investigation of multispecies biofilm formed by Pseudomonas...compounds on these proteins will be tested. Investigating changes in neuronal activity after ischemic ...Armenia Gohar Tsakanova is a Senior Researcher who investigates the molecular and cellular etiopathomechanisms...

...conformational changes in proteins? How can information propagate through a structured material over a long distance...amplified tens or hundreds of thousands of times by propagating through an enormous lattice of identical subunits...

...False positives in original studies and false negatives in replications are unavoidable, but can be minimized...and hypothesis generation, in which data are investigated to look for unexpected trends or differences...possible null is really a true null and not a false negative. Therefore, they will place special emphasis on...

...challenging. How do you prepare for this shift and navigate the pitfalls? Here's what you need to know.   ...lead to frustration. Clinging to technical tasksDelegation can feel unnatural for those accustomed to ...Type Indicator or DISC assessments can help you navigate team dynamics effectively and foster a cohesive...

...addition, we worked with many collaborators to aggregate a sequence-confirmed series of clones corresponding...part of our Genetic Pathway Reference Set. These Gateway-compatible entry clones are the product of many...described in this paper. Systematic Functional Interrogation of Rare Cancer Variants Identifies Oncogenic...

...and the subsequent product is then digested and ligated into an appropriate vector. Gibson Assembly, GeneArt...maintained. Recombination efficiency is high enough to negate the need for selection usually required in recombineering... Plasmid Cloning Techniques like SLIC and Golden Gate Cloning Learn How to Verify Your Plasmid Read our...

...pigment precipitation, enhances the blue color of negative colonies, and allows for better differentiation...also lead to a white colony. And also false negatives: These are rare, but if a small fragment is inserted...example is the ccdB gene system commonly used in Gateway® Cloning. There are also methods to select for ...

...introduced the “2in1” vector cloning system - a Gateway compatible approach to simultaneously clone two...Grefen and Blatt, 2012). Similar to traditional Gateway™ cloning, the GOI first needs to be cloned into... the GOIs into the 2in1 vector instead of the GatewayTM cloning BP reaction, where insertion of the GOI...

...geographical regions When we originally started gathering this information, many assumed that those living...location of the institution. Continuing efforts to gather information and expedite change We are continuing...advocating for and with junior scientists, to help them navigate the scientific enterprise and promote their voices...