...Marcotte EM, Voigt CA. Nature. 2005 Nov 24;438(7067):441-2. PubMed. Find the plasmids from this publication ...Levskaya A, Voigt CA. J Mol Biol. 2011 Jan 14;405(2):315-24. doi: 10.1016/j.jmb.2010.10.038. Epub 2010...fact, precise light-based control of biological processes is highly useful in other fields, including synthetic...to directly activate and inactivate biological processes. Chemical effectors are suboptimal for a number...intensity and wavelength can be used to study dynamic processes. Conversely, varied input patterns may also be...

.... Let's look at an experimental example in Figure 2. Figure 2A shows the sample prior to compensation.... line with the other populations.   Figure 2: Compensation example based on experimental sample..., you will see the populations move, as in Figure 2, following compensation. Figure 3: Flow plot...In flow cytometry, compensation is the process of correcting spillover from one fluorescent channel to...

...design primers to detect the presence of your insert; 2) set up a standard PCR reaction (primers, dNTPs, polymerase...strategies for primer design: 1) insert-specific primers, 2) backbone-specific primers, and 3) orientation-specific...negative clone (-) will not. Backbone-specific primers (2) give larger sized products for positive clones (+...sequencing to verify the insert sequence before proceeding with your experiment. Shorter amplicons tend...

...specific control of activity. Read More CRISPR Class 2 C lustered R egularly I nterspaced S hort P alindromic...frequently neurons) and their governing biomolecular processes. The tools and technologies developed for optogenetics...

...the system outlined in Schmid-Burgk, J.L., et al (2). In this post, I’ll cover two great applications ...molecularly evolved from mNeptune2 through 33 mutations and 2 deletions. CyOFP1 has a higher quantum yield, brightness...luciferin 24 hours later demonstrated a more than 2-fold brighter signal from Antares as compared to Fluc...PMID: 22894855. PubMed Central PMCID: PMC3501149. 2. Schmid, J.L., et al. (2016) CRISPaint allows modular...Here, NLuc internally excites the fluor through a process known as bioluminescent resonance energy transfer...

... intensity of the cell’s illumination (see Figure 2). H describes the maximum signal strength, while W...SSC-A, so most people use that setting. Figure 2: As the cell passes through the laser beam and causes...scale in proportion (while H and W do not; see Figure 2), single cell events will show up in a straight line...heterogeneous cell population, for example from a processed piece of tissue, where cells are present that ...structural complexity. After several steps of tissue processing prior to running my flow analysis, I can however...interest). Let’s double-click inside our gate and proceed to the next plot! Pro tip! If you only get very...

...guides that had zero mismatch off-targets. Guides with 2-bp mismatch off-targets were only toxic if they had...tolerated than mismatches more distal to the PAM, and 2) guides with high GC content have greater off-target...PMID: 28474669. Pubmed Central PMCID: PMC5424143. 2. Tsai, Shengdar Q., et al. "GUIDE-seq enables genome-wide...for identifying genes involved in a biological process. These screens provide an alternative to genome-wide...

...properly exiting the ER and fluorescing (see Figure 2) [5].  When using FPs containing two cysteine residues...compartments including the secretory pathway (Figure 2), the inner membrane space of mitochondria and chloroplasts...PMID: 18228466. Pubmed Central PMCID: PMC2875081. 2. Snapp, Erik Lee. "Fluorescent proteins: a cell biologist's...dark fusions is likely misfolded, blocked from proceeding through the secretory pathway, and retained in...

...while increasing the standard mutation rate by only ~2-fold over nCas9 levels. Two sets of modifications ...10-3 mutations per nucleotide per generation, and (2) the addition of the thioredoxin-binding domain (TBD... the nick site, which was expected since the processivity window of PolI is 15-20 bp. EvolvR’s on-target...bacteriophage T7 DNA polymerase increased the processivity of PolI3M, and thereby increasing the editing...

...The groups in these programs typically consist of 1-2 mentors and 3-5 mentees who meet together for an 8... together. If you all work at the same place, a 1-2 hour lunch on a regular schedule might work. If not...easier with more perspectives contributing to the process Senior people (of which there are never enough,...during the meeting.  Scientists do better with process, so don’t just get together and chat. Have an agenda...started. Have regular meetings. Experiment with the process. Have each person write a Development Plan. Set...

... to share information about the emerging SARS-CoV-2 virus quickly and openly. By sharing results faster...Open Scientific Communication Ecosystem Fig. 2: The scientific ecosystem, enriched by open access...These each have different goals, formats, and processes, and they provide different amounts of access ...journal-based peer review is usually a confidential process among a few individuals.  Early-career researchers...errors, with a lack of transparency in most review processes. Preprints and their feedback, in contrast, are...

... (1) the struggle between two opposing forces or (2) the result of competing desires or (3) the presence... an introduction, and a resolution.   Fig. 2: The structure of a story - not just for fiction! ...find it within the science itself - within that process of failing, over and over again until you don't...

...recognize Fab fragments from the same species and 2) these fragments will be more likely to bind (and ...epitopes recognized by the secondary.   Figure 2: There are a few common solutions to avoid background...Created with biorender.com. Proper tissue processing and sample preparation can also help reduce the...

...panning and scFv characterization.    Figure 2: Phage display starts with a pooled library of scFvs...the short epitope GCN4, also called a scaffold; and 2) a GCN4 binding scFv that’s fused to GFP. When the...Fv (Variable Domain) (scFv) Fragments. Antibodies 2:193–208 . https://doi.org/10.3390/antib2020193  Khantasup...is displayed on the coats of bacteriophages, a process which links genotype and phenotype. Incubating ...single-chain Fv analogue produced in Escherichia coli. Proceedings of the National Academy of Sciences 85:5879–5883...

...nonsense-mediated decay. Although Nelson et al. observed only 2% genome editing in one experiment, they found the ... cells throughout the body, especially stem cells 2. Remove any immunogenicity of the AAV vector Once ...Science. 2015 Dec 31. doi:10.1126/science.aae0169 2. Long C, Amoasii L, Mireault AA, McAnally JR, Li H... mature tissues - no problem, as exon skipping proceeds through the non-homologous end joining (NHEJ) ...is upregulating HDR in mature tissues, as this process is necessary to precisely edit point mutations....

...11 and Cas13 are 1). non-specific RNA cleavage and 2). cell toxicity. Cas7-11 does not display collateral...Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Cell, 163(3): 759-771 (2015). 10.1016...complementary site in the genome where it will proceed to generate a blunt DNA double strand break. Cas9...stranded DNA degradation. Cas3 DNA digestion can proceed up to multiple kilobases, but only in one direction...) and does not require a tracrRNA (it can self-process pre-crRNA into crRNA). The size and simplicity ...11 cuts ssRNA and is capable of pre-crRNA self-processing. Functionally, it is very similar to Cas13 and...

...the nuclear export and translation processes as well.   Figure 2: The conserved eukaryotic polyadenylation...due to the presence of additional helper sequences (2-3). As alluded to above, termination and polyadenylation...transcriptional unit (such as a gene) and initiate the process of releasing the newly synthesized RNA from the...terminator also plays an important role in RNA processing and contributes to variability in RNA half-life...Although mostly thought of as a eukaryotic-specific process, prokaryotes also add poly(A) tails to certain ...studied as prokaryotic termination, the basic processes for eukaryotic termination are understood and ...to terminate transcription. Polymerase I uses a process similar to the prokaryotic Rho-dependent mechanism...

...be decorated with proteins and remain functional, 2) break membranes outside of their natural context ...PubMed PMID: 3759909. PubMed Central PMCID: PMC213417. 2. Schrallhammer, M. et al. Tracing the role of R-bodies...the importance of understanding these complex processes can’t be overstated, we can also learn a lot from...

...parts: (1) β-strands 1-9 (β1-9) and the one ɑ-helix, (2) β-strand 10 (β10), and (3) β-strand 11 (β11). Each...activating caspase-3, and detected fluorescence within 2 to 5 hours, a time consistent with previous methods...plays a pivotal role in an array of biological processes including development, the immune system, and ...cell turnover. Apoptosis is a highly controlled process that is triggered by internal and external signals...

...represent the preexisting resistant subpopulation 2. Resistant cells among the separate replicates would...medicine 21.5 (2015): 440-448. Pubmed PMID: 25849130. 2. Hata, Aaron N., et al. "Tumor cells can follow distinct... she could “get exposure to the drug discovery process.” The ClonTracer library originally came about ...that these are selected for during the treatment process. The authors went on to show that combination treatment...