...strain (which will be covered in more detail in part 2 of this blog series) controls the cell types in which... the nucleotide sequence during the DNA repair process. The addition of small molecules that promote homologous...Also, because non-homologous repair is a random process, zygotes edited using the same nuclease may not...

...aspects of the work you presented may also be unclear. 2. Make the audience your primary focus When writing...and engage with the public, and learn from the process of writing papers and proposals.  To get the most...

...axon terminals to the soma (retrograde tracer) (Fig. 2). Viruses as neuronal scalpels Viruses can be injected...way the brain connects across these regions, a process driven by neurons. By studying how neurons are ...

...to visualize multiprotein complexes.  Fig. 2. Multiplex immunolabeling with subclass-switched recombinant... methodologies drew me to learn more about the process that was used to create this important collection...

...the necessary steps to perform:      Fig. 2: Quick-stepping to a FASTA file   With our outline...output from the first process to the second process. For example, our bbduk process created trimmed FASTQ...pipeline, we wrote a separate Nextflow process. In Nextflow, a process defines which command or script will...specifying the output of an initial process as the input to a subsequent process, we linked each step in our ...pipeline steps into processes and then connect the steps using the output from one process as input to the... Nextflow process.  Next, we specified the input and output channels for the initial process and tested...was clear: to build one process at a time and connect each subsequent process to the existing ones. Once...

...that numbers are not switched around or misread.  2. Track the container - It is just as important to ... be tracked easily at each step in the storage process. After the original sample is barcoded, the lab...stocks, prepped DNA ready for our quality control processes - all of these samples have the 2D barcodes mentioned...a rack of tubes is dropped at any point in the process, we know exactly which location each should go ...step in the receiving, transforming, and storage process, we ensure that the sample tubes that we use match...Addgene’s secret to success. It helps us quickly process samples accurately and in a way that can be easily...deposited samples along every step in the storage process - from deposited DNA to transformed cells and finally...

...experience and to build upon your existing skills.  2. Find your niche Identify and develop an area of expertise...the grant development and management process or the whole process, including grant readiness, funding ... that likelihood. We provide expertise on the processes needed to successfully implement, manage and close-out... funds available and the increasing regulatory process for grants has fostered the demand for experts ...position that required some involvement in the grants process and managing programs that were grant funded. I...and experience through the candidate application process, the candidate’s ability to write a grant application...

...externally fertilized, transparent progeny (Figure 2). These features give it a leg up (or a fin up?) over...powerful resources for investigating any biological process. High quality genome sequence A high-quality sequence...understanding the gene networks that regulate processes from development to metabolism to fertility. Several...mutations in genes affecting diverse biological processes—not just development—are available from strain...Compared to the opaque mouse embryo, where similar processes occur over days inside the mother, the zebrafish...and explain how signaling pathways control this process. Additional zebrafish applications and conclusions... providing unprecedented views into cognitive processes like decision making (Kim et al, 2017). “Brainbow...

...unc- 119 + selection." MicroPublication 1 (2018): 2–3. Wang, Han, et al. "cGAL, a temperature-robust GAL4...interference (DIC) microscopy and computer imaging processing. The combination of these methods provides unique...transgene control in Caenorhabditis elegans." Proceedings of the National Academy of Sciences 115.15 (2018...

...IgM, IgD, and IgG are deleted.     Figure 2: Isotype switching requires DNA recombination of the...next to a different constant domain gene. This process is called isotype switching and results in the ...

...state-of-the-art methods of sex and gender analysis and 2) provides case studies with concrete examples of how...sex-based influences on physiologic or pathologic processes. Much progress has been made in this area in recent...

...that view (Callahan et al., 2022).     Fig. 2: Students in an MDH CURE replaced select residues ...purify, and analyze MDH in semester one and then proceed to use those skills to design and carry out their... become more actively involved in the learning process. As trainees become more comfortable with the open-ended...encouraging them to engage in the problem-solving process. His previous experiences as a jazz musician, serviceman...of students active participants in the research process – so if this sounds like an exciting opportunity...Mentorship and protégé success in STEM fields. Proceedings of the National Academy of Sciences, U.S.A. 117...

...GUS-staining is a very useful visual reporter (Figure 2). The GUS enzyme cleaves a substrate, which will then...plants doesn’t always have to be a time-consuming process. While developing transgenic hairy root lines in... longer, a transient plant leaf transformation process could save the plant biologist some time… months...This method is commonly used in C. roseus, but processes specific to photosynthetic active tissues cannot... In the laboratory we can utilize this natural process by replacing the genes on the T-DNA region with...other aspects of the transient transformation procedure (Mortensen et al., 2019). Our method might provide...

...You’ve probably heard that only 2% of our genome is made of protein-coding genes, and you might be wondering...important roles in many cellular and regulatory processes, but much of their functionality is yet to be ... of microRNA lead to its depletion and Drosha processing retardance. RNA Biol. 2014;11(10):1243-1249. ...

...system (by measuring the fluorescence intensity); and 2) to visualize the localization of the FP (fused to...Therefore, these can be used as “timers” for cellular processes following their activation. The four main FTs ...Throwing color lights on molecular and cellular processes” Curr. Protein. Pept. Sci. 9(4):338. Chudakov ...

...replicate, then purify the DNA.     Figure 2: A pooled library page with protocols boxed in red...!     Figure 1: Flowchart showing the process for using pooled lentiviral libraries. A) Amplification...of your pooled library during the amplification process. It’s far better to verify via NGS now than to ...want to consider spinfection. Spinfection is the process of transducing cells at a concentrated seeding ...

...mazF gene and a gene for Zeocin resistance (Figure 2). This MazF-ZeoR cassette is flanked by two direct... toxin which can affect a variety of cellular processes and an antitoxin that suppresses the toxin’s activity...host cell division to plasmid proliferation." Proceedings of the National Academy of Sciences 80.15 (1983... model for programmed bacterial cell death." Proceedings of the National Academy of Sciences 93.12 (1996...

...PMID: 18478103. PubMed Central PMCID: PMC2364656. 2. Dittmar, Kimberly A., et al. "Selective charging ...need fewer resources for the entire translation process. For example, during high growth rate conditions...segregate with the cell pellet during purification procedures. Even if you produce a large amount of protein...determined by both codon bias and folding energy." Proceedings of the National Academy of Sciences 107.8 (2010...

...156–159. https://doi.org/10.1016/0003-2697(87)90021-2 Chomczynski P, Sacchi N (2006) The single-step method... products that help to further streamline this process and/or stabilize RNA during tissue collection and...

...1) or added using PCR-based amplification (Option 2). Advantages of Golden Gate cloning Golden Gate cloning...enzyme recognition sites. As a result, the ligation process is close to 100% efficient. Another strength of...

... adapted to a competitive format. Figure 2: Overview of using Competitive ELISA to quantitatively...pair. In a sandwich ELISA, the first step of this process is the addition of a detection antibody (typically...

...protocols 4.4 (2009): 495-505. PubMed PMID: 19300443. 2. Geraerts, Martine, et al. "Comparison of lentiviral...subcloning, expansion, and confirmation. This procedure can take several months but should be considered...

...to the break room for a sip of water.  Fig 2: Blugene likes to spark creativity in out on the open...because a process is working, doesn’t mean it will always be that way. Things change and processes often ...intentionally engage in a creative problem-solving process?  You may want to start by carving out time to ...holds the dual purpose of giving people time to process the issue properly while also discouraging everyone...is thrown out because they haven’t had time to process on their own. You may also find your meetings run...This is why perfectionists tend to also be procrastinators.  If you fall into this category, get out of...

...systems utilize with type I-F Cascades. Figure 2: Cascade complex from CRISPR type I-E. Created with...components of Cascade for maximum efficiency of the process. Various labs have designed all-in-one Cascade ...needs to remain anchored to Cascade throughout the process. The reeling of ssDNA and unwinding of dsDNA occur...

...PubMed PMID: 3459152. PubMed Central PMCID: PMC323600. 2. Chester, Nicholas, and Daniel R. Marshak. "Dimethyl... mutagenesis vastly facilitates the subsequent process of screening for succesfully mutated clones. Forward... introduce undesired mutations through the PCR process, you need a high-fidelity polymerase. There are...fragment length polymorphism (RFLP) analysis. In this process, if your mutation introduces an additional restriction...DNA duplex stability from the base sequence." Proceedings of the National Academy of Sciences 83.11 (1986...Yongzhen, et al. "New insights into the QuikChangeTM process guide the use of Phusion DNA polymerase for site-directed...

...Nature 339 (1989). 487–488. PubMed PMID: 2725683. 2. Miller, C.J., Kassem H.S., Pepper S.D., Hey Y., Ward...target cells Labs tend to follow the same general procedure for lentiviral infections, produce the virus, ...vectors are engineered with non-native envelopes, a process called pseudotyping. Lentiviruses pseudotyped with...Segura, M.M., Kamen, A., Garnier, A. “Downstream processing of oncoretroviral and lentiviral gene therapy...cellular receptors for vesicular stomatitis virus.” Proceedings of the National Academy of Sciences 110.18 (2013...

...condition.   Fig. 2: Schematic of the goldilocks antibody selection process The exposure time ...

.... Find Plasmids from the publication at Addgene 2. Green, Alexander A., et al. "Toehold switches: de-novo-designed...technology through product development, the regulatory process, and manufacturing so that we can get the diagnostics...

...of 3,5-difluoro-4-hydroxybenzylidene-imidazolinone-2-oxime (DFHO), a fluorophore modified from DsRed. DFHO...Fluorescent RNA aptamers are used to track cellular processes by transforming cells with plasmids encoding aptamer-tagged...for qualitative and quantitative measurement of processes including RNA movement by tagging RNA sequences... Aptamer Fluorophores describes the enrichment process used to evaluate oligonucleotides for binding affinity...affinity to a specific target, and the molecular processes that make aptamer biology a powerful tool. In ...

...communicate the nuances of protein structure in a flat 2-D images. In the early days of structural biology,...protein. At some point during this interrogation process, they will almost invariably say something like...

... already have an end product in mind, steps 1 and 2 will generally be straightforward. Likewise for step...that need to be considered during the planning process. For CRISPR experiments, the following framework...

...PMID: 24901218. PubMed Central PMCID: PMC4047077. 2. Pagliara, Patrizia, et al. "Ostreopsis cf. ovata ...bisphenol A-induced perturbation in biological processes in ER-negative HEK293 cells." PLoS One, 2014. ...

...PMID: 25538079. PubMed Central PMCID: PMC4353587. 2. Stephen, Andrew G., et al. "Dragging ras back in ...from rogue cells when something does go awry. The process of programmed cell death, or apoptosis, is one ...

... Werner S, Marillonnet S. PLoS One. 2011 Feb 18;6(2):e16765. doi: 10.1371/journal.pone.0016765. PubMed...no PCR, no gel purification; the whole cloning process is so simplistic once you have your parts. That...

...variety of environmental conditions, and double every 2 hours. Yeast are also the first eukaryotic genome ...actually share many biological properties and processes with our own cells. Like human cells, yeast DNA...proteins and tissues to better assess developmental processes. For example, the Stainier lab studied the proliferation...making it ideal to study basic molecular biology processes. C. elegans are also quite amenable to genetic...investigate an array of biological systems and processes including the nervous system, development, cell...β-cells identified by HOTcre mediated labeling." Proceedings of the National Academy of Sciences 106.35 (2009... in freely behaving Caenorhabditis elegans." Proceedings of the National Academy of Sciences 113.8 (2016...

...PMID: 27151215. PubMed Central PMCID: PMC4938170. 2. Zuris, John A., et al. "Cationic lipid-mediated delivery... to genome editing of mouse zygotes, through a process known as CRISPR-EZ (CRISPR RNP Electroporation ...

... the Cas machinery where to cut.      Fig. 2: Heatmap indicating loci where SNPs reside on sgRNA...with an ancestry-based bias.   The correction process does not stop there. Currently, they are working...

...of these issues are not particularly new. Chapter 2, “The Professional Scientist,” addresses the challenges...consequence of the undermining of the scientific process and the marginalization of science, particularly...

...community is active, engaged, and has great questions! 2. The r/Science subreddit is well moderated. One of...You can see some of Joanne’s comments on the AMA process below, but we also wanted to share some thoughts...submission to avoid rousing the trolls. 3. The AMA process was simple and well organized. You can check out...

... Methods 2009; 6, 343-345. PubMed PMID: 19363495. 2. Wang JW, Wang A, Li K, Wang B, Jin S, Reiser M, Lockey... menu of common cloning options in the deposit process in 2014 because of its gain in popularity. Gibson... the Miller Lab Protocol). The Gibson assembly process can be used to assemble up to 6 fragments in one...time commitment. Another advantage is that this process makes it easy to generate wild type and mutant ...drawback to the Gibson assembly technique is that the process works best with fragments over 200 nucleotides....

...rules are: (1) don’t use the same overhang twice; (2) avoid palindromes; (3) no overhangs with the same...genomes, it required a very stringent selection process and benefited from a longer reaction time of 15...

...al (Nature. 1988 Jul 28;334(6180):320-5.) PubMed. 2. A segment of the 5' nontranslated region of encephalomyocarditis... a novel gene, is not always a straightforward process. Rather than try to directly detect your gene of...

... vivo and in vitro applications.   Figure 2: Modular features of DIRECTED particles where the ... more accessible by simplifying the production process and could even enable the creation of CAR-T cells...

...rate of 10-55% - the PASTE system.     Fig. 2: Schematic of programmable gene insertion with PASTE... a guide RNA. The enzyme has dual activities: processing pre-CRISPR RNA into mature CRISPR RNA and cleaving...

...Nutr, 129 (2009a): 477–484. PubMed PMID: 10064313. 2. Chapman-Smith, A., & Cronan, J. E. “The enzymatic...These biotin dependent enzymes are essential for processes such as gluconeogenesis, lipogenesis, amino acid...energy transduction. Biotinylation is the general process by which biotin is added to a protein or macromolecule...biotinylation occurs naturally in cells and is the process by which biotin is covalently bonded to a specific...

...become a valuable asset yourself.     Fig 2: A diagram of the Thomas-Kilmann Conflict Model, courtesy...communicated. The next step is to come up with a viable process that will avoid such miscommunications in the future...and examples that have guided your resolution processes in the past. Types of Resolution There are of...

...gene1 [lon] ompT gal sulA11 R(mcr-73::miniTn10--TetS)2 [dcm] R(zgb-210::Tn10--TetS) General protein expression...auxillary plasmids may be isolated during the prepping process. How does inducible expression work? As mentioned...

...Postdoctoral Associations around Boston - From Panel Session 2: Defining the Postdoc: An Institutional Perspective...questions or concerns throughout the publishing process. The diversity panel - Equity and inclusion The...

...signal separately for each target.   Figure 2: Examples of simultaneous multiplex IF labeling of... appropriate secondary antibodies. The general process is similar to standard immunofluorescence, but ...spatial analysis of cells in complex tissues. Proceedings of the National Academy of Sciences, 117(52),...

...of concept for further development. Figure 2: Mechanisms of deaminase-based adenine transversion...editor. Black arrows indicate natural cellular processes. Created with BioRender.com. The first deaminase-free...