...detection kits that use detector cell lines, ELISA, fluorometry, or PCR-based approaches to detect even low levels... storage buffers used, and other experimental parameters. For example, while freeze thaw is generally ...H.G. “Detection of mycoplasma contaminations.” Methods in Molecular Biology 946 (2013). 1-13. Pubmed PMID...PubMed Central PMCID: PMC3456965. 11. Zhang B., Metharom P., Jullie H., Ellem K.A., Cleghorn G., West M.J...

...years, but it usually ranges from 20-50 years and sometimes longer. There is no known safe level of exposure...has caught it has usually started to spread (metastasize) and turned into Stage 3 or Stage 4 cancer. These...chemotherapy drugs used to treat mesothelioma – pemetrexed and cisplatin –because it cuts off the blood ...in the world. The development of new diagnostic methods and treatments such as these has brought a lot ...

...perturbing circuits together with the development of methodologies to deliver genes to the CNS have recently made...use epitope tags to monitor single molecules - something researchers often want to do at neuronal synapses...microscopy. Sample preparation for these techniques sometimes makes it more difficult to detect protein tags...probes for light and electron microscopy." Nature methods 12.6 (2015): 568. PubMed PMID: 25915120. PubMed...

... for your brainstorm session is to start with something completely unrelated to the issue and consider...out-of-the-box concept and pare it down until you have something tangible and applicable to your situation.   Thinking... We become so fixated with the idea of making something the best it can be that it seems impossible to...stuck on a problem is to step away from it. Find something unrelated to do (many Addgenies enjoy rock climbing...

...understood, but some possible reasons include: Metabolic pressures - it takes cellular resources to produce...optimized for a particular organism, this will sometimes (but not always) be noted in in the “mutation”...of your protein of interest. Regardless of which method you choose to overcome the problems surrounding...surrounding codon choice, you should have some method to make sure the proteins you produce function properly. ...

...pair palindromic repeats separated by an 8 bp asymmetric core. Recombination between these sites can lead...discovered that it is not the exact sequence of the asymmetric target site core but its 8 bp length that is ... recombinase activity (Logie and Stewart 1995; Metzger et al. 1995). Visualizing single neurons in the...PMID: 7597057. PubMed Central PMCID: PMC41617. 7. Metzger, D, J Clifford, H Chiba, and P Chambon. 1995. “...

..., but for many of us, scientist is something we are, not something we do. We have to find a way to keep...try to keep discussions on-topic and, at least sometimes, civil. r/Science in particular hosts a variety...scientists in training is abundant, helpful, and sometimes excellent. Finally, as I said in the AMA, it’s...

... restriction/methylation system. This means the strain can neither restrict nor methylate DNA. dcm: Similarly...Similarly, strains with this mutation are unable to methylate cytosine within a particular sequence. Browse...of T7 RNAP from the genome; does not restrict methylated DNA fhuA2 lacZ::T7 gene1 [lon] ompT gal sulA11...

...Journal of Neuroscience Methods, 172(2), 250–254. https://doi.org/10.1016/j.jneumeth.2008.05.003 Ghosh, R...concentration changes in response to experimental parameters. This is a job for a semi-quantitative western...not especially linear. In panel C, two different methods of quantifying total protein are compared. While...

...helpful. I set out to make something practical and accessible. If there's something you come across that is...The second question is whether there is chalk. Sometimes. It can also be a slide deck, markers, etc. Chalk... What is the big question? This should be something that anyone could relate to, that people would...

...encoded sensors enable real-time observation of metabolite production." Proceedings of the National Academy... has recently been used to study a matrix metalloproteinase: López-Pelegrín, Mar, et al. "A novel mechanism...mechanism of latency in matrix metalloproteinases." Journal of Biological Chemistry 290.8 (2015): 4728-...Christopher Marx's lab has engineered and validated a new method called FREQ-Seq. This strategy allows scientists...

...resembles a cylinder with a length of 4.2 nm and a diameter of about 2.4 nm (Hink et al., 2000). The complete...Quantitative assessment of fluorescent proteins." Nature methods 13.7 (2016): 557. PubMed PMID: 27240257. PubMed... expressed inositide binding protein domains." Methods 46.3 (2008): 167-176. PubMed PMID: 18930153. PubMed...proteins to study mRNA trafficking in living cells." Methods in cell biology 85 (2008): 273-292. PubMed PMID...

...cassette. This leaves a “scar” in the chromosome, sometimes interfering with expression of surrounding genes...leave any undesired scars in the chromosome. These methods roughly fall into two categories: those that rely... a sucrase that converts sucrose into a toxic metabolite. Cells that have undergone a double crossover...identify double crossovers. This can be an efficient method to generate precise, unmarked mutants, but it frequently...

...approach combines the overlapping and trans-splicing methods. The AAV encoding part A of the transgene contains...of the transgene have been seen with the hybrid method. While these fragments are likely non-functional...Frischmeyer et al., 2002). How to pick a split AAV method While the hybrid dual vector system has a slight...Adeno-Associated Virus Capsids. Human Gene Therapy Methods 27:1–12 . https://doi.org/10.1089/hgtb.2015.140...

...serotypes have a primary cellular receptor and sometimes one or multiple coreceptors, which may be present...present on different cell types and sometimes vary between target species or even strains. Pro tip! Getting...underexplored sites on the AAV capsid. Molecular Therapy. Methods & Clinical Development, 26, 331–342. https://doi.org...2011). AAV-mediated liver-directed gene therapy. Methods in Molecular Biology (Clifton, N.J.), 807, 141–...

...failure and problem solving. I believe that is something lacking during degree courses; they never really...the company was to fulfill contracts, my work sometimes felt rushed and the focus always seemed more on...situation, communication between departments was sometimes difficult and the solution attempted was more ...solved with good management practices, but can sometimes go unnoticed at large organizations. Another downside...

...Surprisingly, the team noticed that this editor sometimes introduced C → G edits at certain positions within...multi-step cellular repair pathway that is known for sometimes causing double-strand breaks (DSBs). The Fei-Long...common intermediate of CGBE activity, and they are sometimes processed by error-prone non-homologous end joining...context preference predictable by machine learning methods. Nature Communications, 12(1), 4902. https://doi.org...

...and methods section. This section will vary slightly depending on the application — flow cytometry, western...  Figure 4: A screenshot of the materials and methods section for the data report shown in Figure 3. ...

... database. Previously developed genome editing methods using nucleases and base editors have the potential...less constrained by PAM availability than other methods. The PAM-to-edit distance can be over 30 base pairs...can be used with any of the PE2-5 approaches, sometimes referred to as PE2max, PE3max, etc. PE6: Specialized.... Although the addition of the RNA pseudoknot sometimes altered epegRNA expression compared to pegRNAs...

...mechanism of matrix metalloproteinase-9 secretion, and identifying lysine methyltransferase substrates. We’...genome-wide libraries for CRISPR screening." Nature methods 11.8 (2014): 783-784. PubMed PMID: 25075903. PubMed...