...research project must have innovative yet attainable milestones that contribute to intellectual thought and discovery... and skills do you need to use to reach these milestones? Can you get assistance from people who have ...Think about what your personal goals are and set milestones along the way so that you don't get stuck doing...been developed. Can you think of a project that combines an exciting new method or approach with another... Passion fuels the drive to continue persuing milestones and working hard. Being patient, being selective...

...validated simultaneously in the brain and the intestines of mice. The result? A very cool tool to advance...polymers are heated up and pulled to a desired thickness. The fibers are drawn with microelectronics included...engineering road map of sorts, giving an idea of the stiffness the fibers would need to be, as well as other ...fiber implants. Due to the soft nature of the intestines and the requirement for digestive motility to...excited a class of enteroendocrine cells within the intestines known as neuropods. They observed increased vagus...

...Now that we know all about antibiotic resistance genes, let’s consider another basic element of any plasmid...focused only on those found in bacteria. Some common ones you might see include ColE1, pMB1 (which comes in...20-40 CloDF13 (CDF) D Relaxed This table defines common cloning vectors, their copy number, ori,...copies of plasmids if they contain large inserts or genes that create a toxic product. The E. coli strain...for single- and multiple-unnatural amino acid mutagenesis in Escherichia coli. Biochemistry. 2013 Mar 12...

.... Yet, we cannot prosper in a world where our medicines, technologies, defense, and governing policies...more scientists in our leadership, and raising awareness regarding the pitfalls of accepting alternative...chromosomes. Montreal, QC, Canada 45.5017°N 73.5673°W – Vanessa Sung Hundreds of scientists and supporters in ...well as allies outside the scientific community. Vanessa Sung is currently a PhD student studying breast...restorative justice.  Follow her on twitter @sung_vanessa. Albany, NY, USA 42.6526°N 73.7562°W – Nicholas...denying science, from celebrities falsely claiming vaccines cause autism to politicians saying that climate...

...mice Transgenic mouse lines have also been engineered to express various opsin genes [7-9]. The advantage...other tool in a scientist’s vast toolbox, its effectiveness depends on how and why it’s used. There are ...behavioral output is sufficient to assess the effectiveness of your optogenetic manipulations (though a ...has genetically manipulated a number of the opsin genes to alter the light spectrum for activation and the...advantage of using these lines is, if one is crossed with an appropriate mouse line expressing a particular ...

...Julio Ernesto Valdivia Silva, Universidad de Ingeniería y Tecnología (UTEC), Peru Julio Ernesto Valdivia...her current research, Sheen aims to identify new genes related to PZA-resistance by using different approaches...experience in the field of bacterial and fungal pathogenesis. Her research focuses on a collection of pathogenic...growth will then be measured to link the deleted genes to specific functions in biofilm formation. The...

...Cecchetelli Auxin Inducible Degradation (AID) uses plant hormones called auxins to conditionally and reversibly ...AID to yeast, as well as mammalian and human cell lines (Nishimura et al., 2009, Natsume et al., 2016).  ...protein of interest to create conditional human cell lines. Lastly, the Kanemaki lab demonstrated that auxinole...targeted by the Cas9 nuclease. The Streptococcus pyogenes Cas9 (SpCas9) requires an NGG PAM to recognize... role in the transcriptional regulation of many genes. A number of groups have designed means of studying...

...provide the Brunello (targeting human genes) and Brie (targeting mouse genes) pooled libraries from the labs...popular genome editing tool is its developers’ willingness to make their CRISPR technologies available to...pooled gRNA libraries that can be used to screen for genes involved in a wide variety of cellular processes...80,000 gRNAs to target around 20,000 individual genes. We provide enough lentivirus for you to both perform...

...when choosing an individual fluorescent protein (brightness, photostability, and so on; see the previous ...use (particularly not the most recently published ones), they can be a good starting point. In many cases... used filter sets for multicolor imaging include ones designed for CFP, YFP, and RFP or the Sedat Quad...supplement your cells with biliverdin for maximal brightness. In mammalian cells, one of the improved folding...proteins with this filter set and have reported brightness measurements. Here, EGFP outperforms the improved...

...stable cell lines and are not used for general plasmid selection. Instead, reporter genes are often employed...within the cell for effective replication. Cell lines expressing the Epstein–Barr virus (EBV) nuclear ...48 h post transfection when creating stable cell lines. Looking for Mammalian Expression Plasmids? Check...Read More: Plasmids 101: Antibiotic Resistance Genes Plasmids 101: Origin of Replication All Plasmids...

...global pandemic in which scientists race to find vaccines, therapeutics, and other treatments to stem the...plasmids containing all SARS-CoV-2 protein coding genes in E. coli and S. cerevisiae expression vectors ...yeast expression constructs contain protein coding genes, tags, truncations, and codon optimizations similar...plasmids. Each combination is present in two sets of backbones which drive expression using either TEF1 or GAL1...of plasmids containing SARS-CoV-2 protein coding genes in E. coli and S. cerevisiae expression vectors ...

...transcription of other relevant viral genes (including the ones listed previously). E3/E1B suppresses ...applications and aid in ease of use. Our lentiviral guide outlines these components.     Fig. 1 - Viral vector... AAV also requires some back up from adenovirus genes – E4, E2a, and VA – which are supplied on a helper...viral replication. More information on these viral genes is available in the adenovirus section below!   ...

... comic, when a bacterium uptakes a plasmid, the genes on the plasmid can give them different phenotypes...example are bacteriophages that attach to bacterial membranes and inject their genetic material into the cell... DNA into eukaryotic cells, like mammalian cell lines. This can be done with lentiviral and Adeno Associated...and AAV can be used to create both transient cell lines, where a gene of interest is expressed but not integrated... integrated into the genome and stable cell lines, where foreign DNA is incorporated into the cell’s genome...

...activated by an array of different ligands, including hormones, neurotransmitters, photons or odorants, to elicit...that there are approximately 950 different human genes encoding GPCRs (Takeda et al., 2002). G proteins...be used to detect G protein activation via bioluminescence resonance energy transfer 2 (BRET2). Check ..., GPCRs were first mutated via site-directed mutagenesis to bind non natural ligands. These non natural...2002) Identification of G protein-coupled receptor genes from the human genome sequence. FEBS Letters 520...

...single-use” transgenic lines. Instead, researchers can generate and maintain transgenic lines that can be used...system that makes use of pre-existing transgenic lines, termed BLITZ (Biotin Labeling In Transgenic Zebrafish...dCas9 and microbial SSAPs (dCas9-SSAP) which insert genes in a cleavage free manner. The dCas9-SSAP system... 12-plex multiplex imaging system. Mixing the subclones within a single well reduced well-to-well variability...

... nucleic acid sequence of the antibody-encoding genes over time as the hybridoma cells divide. These changes...hybridoma cells before culturing or they can clone the genes from the hybridoma into a plasmid to create a recombinant...Figure 2: Changes to the heavy chain or light chain genes within a hybridoma impacts the specificity, avidity...Wilkinson IC, Tandon AK, Anderson S, Bladen CL, Jones B, Aldred SF, Bestagno M, Burrone O, Maynard J, ...1999. 1, Generation of Hybridomas: Permanent Cell Lines Secreting Monoclonal Antibodies. https://www.ncbi.nlm.nih.gov...

...target gene, it therefore determines the timing of gene expression and largely defines the amount of recombinant... cAMP and represses pBad, decreasing promoter leakiness.   Other examples of chemically induced promoters... be compatible with your experimental system. Leakiness If you’re expressing a gene that may be toxic,...inducible promoters are also known for their low leakiness. Inducibility level How much do you want to induce...

...newly synthesized product from a site-directed mutagenesis experiment? The answer to both questions is the...endonucleases. A given prokaryote typically has genes encoding one or a few restriction modification systems...cleave DNA. DpnI is often used for site directed mutagenesis. During this process, incorporation of a desired...coli strain and will therefore have methylated adenines in any GATC sequences found in the plasmid. When...Troubleshoot Your Sequencing Results Use REPLACR Mutagenesis to Mutate Your Plasmids Resources on Addgene.org...

...TRIM21 toxicity, Clift et al. created stable cell lines overexpressing TRIM21, finding that TRIM21 overexpression...contain changes in expression of TRIM21-associated genes, indicating that TRIM21 overexpression does not ... within 4 hr of treatment. In some primary cell lines, endogenous TRIM21 is sufficient for TRIM-Away, ...Recruitment for Epigenome editing by Cas9 (FIRE–Cas9), combines these approaches to enable rapid and reversible...that can independently control expression of your genes of interest. These transcription factors consist...

... al. "High-Resolution CRISPR Screens Reveal Fitness Genes and Genotype-Specific Cancer Liabilities." Cell...et al., 2014; Konermann et al., 2014) individual genes for studies interrogating their functions. The continual...list of effective gRNA sequences targeting your genes of interest. Thanks to the tremendous efforts made...Hongming, et al. "A CRISPR-based screen identifies genes essential for West-Nile-virus-induced cell death...