...“no.” When there was room for misinterpretation, that misinterpretation was definitely happening. Our ...terminology of a new field. Annotate: When creating important documentation, including SOPs or project-...were leaving these meetings with different interpretations of what we discussed and decided. I needed ...definitions in our lexicon. We created documents and presentations, gave real life examples of when and how to ...frequently used words like this when starting rotations or my postdoc would have simplified the first ... ambiguous terms. Defining words and applying annotation can be particularly useful in academic labs when...

... screen plasmids for the appropriate mutations.  Point mutations and other subtle changes should be confirmed...molecular cloning; however, one of its biggest limitations is that sequence modifications can only be made...selectable and non-selectable sequences, point mutations or other small base pair changes, and the addition...nucleotides, while ssDNA substrate is best for point mutations or changes of only a few base pairs.  dsDNA Substrate...the two options, but these cells are prone to mutations and will have more unintended changes to their...Taq polymerase to generate insert ssDNA Point mutations or changing a few nucleotides 0.1-1% or 25-50%...effects of Gam expression and to limit spontaneous mutations that occur when Red is constitutively expressed...

...projects in introductory labs. Genome projects use computational methods to identify genes by their similarities...proteins have acquired divergent functions due to mutations accumulated over time? Undergraduates can help...inexpensive. In addition, a complete set of well-annotated deletion strains is available (6). Methionine ...ura3 mutants and cannot grow without uracil supplementation (6). Expression of MET/Met genes from the pYES2.1...blotting in other experiments (Figure 2).   Complementation reveals conservation of function Students have...Addgene, to successfully analyze Met protein complementation. In the procedure diagrammed in Figure 3, students...promoter controls MET/Met gene expression, complementation should only occur when galactose replaces glucose...

...type of experiment is called a protein fragmentation complementation assay (PCA). In the Split-BioID method...FRET is another example of a protein fragmentation complementation assay that can be used to study binary...individual protein. The Bethune Lab has overcome this limitation by creating Split-BioID, a spatiotemporally controllable...Protein Interactions. Split-BioID is a protein complementation assay where BirA*, an E. coli biotin ligase...proteins. One caveat of Split-BioID is that the orientation of the BirA* fusion matters! Schopp et al saw...interact with the bait proteins. Another potential limitation of Split-BioID is its speed of labeling. BioID...

...number of citations highly influenced by this paper. Influence is measured by the number of citations and the...extract important features such as influential citations, images, and key phrases. It also takes into account... account how papers are connected via citations, in order to return results that are more relevant to ...with greater influence, the average number of citations per year and the number of other manuscripts that... When you view a paper’s entry, the number of citations that paper has is displayed at the top of the ... the context of these citations. The number of manuscripts that cite the background, methods, or results...all publications by an author, total number of citations, and the number of publications that heavily cite...

..., or for generating mutated sequences for site-directed mutagenesis.  Quantitative polymerase chain reaction...genotyping. It’s extremely useful but is not as quantitative as other methods of PCR. Theoretically scientists...reaction, qPCR Quantitative polymerase chain reaction (qPCR) also known as real time PCR is a PCR technique...material in your sample.  qPCR can also be used to quantitate RNA levels using reverse transcription-qPCR (...sequences that are rare, for example, rare alleles or mutations. ddPCR also does not require a reference or standard...is determined and then analyzed using Poisson statistics to determine the concentration of the original...PCRs that use a single pair of primers, like those stated above. Multiplex PCR is also used in high throughput...

...contains a mutation or modification, this should be included in the plasmid name. Mutations are generally...abbreviation of the mutated amino acid currently at that position. In contrast, unmutated or Wild Type (wt...your plasmid name. This is often a 3-6 letter representation of the gene (or DNA sequence). pBACKBONE-hGene...change. The proper way to denote an amino acid mutation is to list the one letter abbreviation of the ...   Example: If the Glutamine at position 295 was mutated to an Alanine, Q295A.  Mutant version: pBACKBONE-Flag-hGene...

...Considering citations to all articles, Anurag Acharya and colleagues report, the percentage of citations to articles...to 1975 and is based on the average number of citations for recent articles. Of course, a publication ...measure scientific impact. Even if we stick to citation counts for a moment, it’s clear that articles ...articles in non-elite journals went from 27% of all citations in 1995 to 47% in 2013 – yes, almost half.   The...Accessibility is surely also behind the increase in citations for older publications. (As an interesting aside...the amount of protein in a solution.) But are citations and journal publications really the best way to...

...COVID-19 cases each week. Statistical Modeling Note -- If statistical modeling isn’t up your alley,... estimated total of 380,000 deaths worldwide (statistics provided by Google on 6/4/20). Many countries... open-ended question is difficult to answer quantitatively without broad, unbiased data. Therefore, we...order several plasmids at once, breaking the statistical independence assumption of the Poisson process...process. This modification also eases the interpretation of the parameters we infer, since scientific productivity...remind the reader that this is not a causal interpretation -- the effect would generally not be the same...His research uses applied machine learning and statistical methods for fundamental scientific discovery ...

...parents looking for experts to come give guest presentations. If you’re a scientist thinking about reaching...already discussed the idea of my giving a CRISPR presentation to their students. I didn’t think of this opportunity...postdocs, and other working scientists, creating a presentation for 8th graders was a little intimidating. Too...received from the teachers, I formulated the presentation as follows (which could be adapted for any topic...do? I spent more time on this section of the presentation because, in my opinion, it was more important...teachers with whom I worked, for feedback on my presentation, she said “it was such a valuable extension ... of CRISPR. Two of the examples I used in my presentation had only become public in the previous two weeks...

... DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) because any engineered DNA-binding...region of the transcription factor IRF-1. Immunoprecipitation, using an antibody targeting the 3xFLAG tag...purification of a specific genomic region, numerous adaptations of this system have been made in different fields...also has great potential for improvements and adaptations that can provide an increased understanding of... DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) using CRISPR." Biochemical and biophysical... DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP)." Scientific reports 3 (2013): 3171...and Hodaka Fujii. "Insertional chromatin immunoprecipitation: a method for isolating specific genomic ...

... an invitation to join the Advisory Board of Addgene, I accepted the offer without any hesitation. I hope...transcription by using locus-specific chromatin immunoprecipitation (locus-specific ChIP). These methods consist...consist of insertional chromatin immunoprecipitation (iChIP) and engineered DNA-binding molecule-mediated...molecule-mediated chromatin immunoprecipitation (enChIP), both developed in the lab. In June 2014, Dr. Fujii joined ..., we developed the insertional chromatin immunoprecipitation (ChIP) (iChIP) technology, which uses locus-tagging... DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP). Fujita T, Asano Y, Ohtsuka J, Takada... DNA-binding molecule-mediated chromatin immunoprecipitation (enChIP) using CRISPR. Fujita T, Fujii H....

...such a small package. But don’t let the simple presentation deceive you; pooled libraries can be quite challenging...verified through NGS sequencing to confirm library representation. Transfection of the DNA pooled library and...transduction of cells, resulting in cell population mutated by the viral pooled library. These cells can then...protocol that amplifies without disrupting plasmid representation. A good place to start is with a protocol for...library by next-generation sequencing (NGS). The representation of your amplified library should be very similar...page, you can find protocols for sequencing, representative NGS results (read counts or another visualization...library and verify that you have not skewed the representation of your pooled library during the amplification...

... an enzymatically “active” state rather than reverting to an inactive state like Cas9 or Cpf1. As a result...proteins like SaCas9 and Cpf1 can circumvent the limitations associated with SpCas9. Cas13a (previously referred... HEPN domains in other proteins. As with Cas9, mutating key residues in the Cas13a molecule results in...system, Cas9 uses both a tracrRNA and crRNA to facilitate binding and cleavage of target DNA respectively...molecule through a uracil-rich stem loop and facilitates target binding and cleavage through a series ... to activate programmed cell death or a dormant state for bacterial cells that have been infected with...Zika virus, genotype human DNA, identify tumor mutations within cell-free genomic DNA, and detect COVID...

...differentiated somatic cell reverts back to a pluripotent state, which then proliferates and is redifferentiated...differentiated cell type to an undifferentiated state and then redifferentiation into the cell type of...Conversion of adult somatic cells into the pluripotent state. Redifferentiation - Conversion of the pluripotent...Dedifferentiation to an intermediate pluripotent state is not always obligatory in cell conversion processes...back to their most primitive pluripotent stem cell state, through transdifferentiation adult somatic cells...factors) More Chances of accumulation of harmful mutations More, due to selection from more cycles of proliferation...objective is to fix a disease-inducing genetic mutation in a patient, trying to transdifferentiate any...

..., I promise it isn’t.) The intuitive, qualitative interpretation of this gel is misleading. We can see...causes misleading qualitative conclusions, and it’s also problematic for computational quantification. ...experimental parameters. This is a job for a semi-quantitative western blot. Most western blots benefit from...Loading controls are even more important for semi-quantitative western blots. No pipetter is perfect, and proteins...lysate (10–40 µg) probed with β-actin (1:2000). Representative of three independent blots. B–C) Graphs showing...high-abundance single-protein controls in semi-quantitative immunoblotting. Journal of Neuroscience Methods...

....1c02201. LiLac sensor for lactate Stop and smell the flowers - or rather, the lactate! The latest biosensor...for lactate developed using a high-throughput multiparameter screen (Koveal et al. 2022). Lactate is a...previous lactate biosensors, and its affinity is better-matched to cellular concentrations of lactate. The...ZAP70 (ZtSH2) which binds only the phosphorylated state of an immunoreceptor tyrosinase-based activation...around 70 mM and 100 mM, respectively. Introducing mutations and using potassium-binding protein homologues...

...bactericidal) or slow growth/prevent cell division (bacteriostatic); however, the distinction between the two ...categories may be a bit of a gray area as some bacteriostatic reagents can kill bacteria when used at high...subunit; inhibits aminoacyl translocation Bacteriostatic 50-100 ug/mL in EtOH Polymyxin B polypeptide...inhibits protein synthesis (elongation step) Bacteriostatic 10 ug/mL Chloramphenicol   Binds 50S ribosomal...ribosomal subunit; inhibits peptidyl translocation Bacteriostatic 5-25 ug/mL in EtOH *In prokaryotes.     ...inactivates the gene and acts as a marker for the mutation. When designing these types of experiments it ...not to use the same resistance cassette for the mutation and for plasmid selection. Additionally, scientists...

...DNA with phi29 polymerase prior to fragmentation can greatly facilitate downstream manipulations. Phi29 ...activity without the need for genome sequencing or annotation (5). In the event that genome sequencing can ...to enzymatic digestion, however, a physical fragmentation strategy is preferable. Nebulization involves...associated with cloning large fragments. After fragmentation, repair sheared ends with T4 DNA polymerase ...gently down onto a plate of colonies. Mark the orientation of each membrane on its plate before moving on...your membrane to several freeze/thaw cycles to facilitate cell lysis: alternating between 5 min at -20°...sequence the inserted DNA and characterize your new putative host cell-binding proteins! Many thanks to our...

...professionals has made possible a significant qualitative and quantitative expansion in the 2015-2016 academic ...does research at UK in the same field that I do: prostate cancer. She has guided me in my next steps. For...baseline self-assessment survey to determine their expectations of the program and their baseline skills. Mentors... as mentors to Spanish mentees, mentors also facilitated the development of bidirectional USA-Europe professional... pre-program self-assessment survey defining expectations of the program and their baseline skills and...improvements in writing fellowships, giving oral presentations, and preparing for job interviews, expanded ...Spanish scientific community overseas. Official representatives from different Spanish universities and high...

...  An MTA is simply a contract or agreement that states the terms of use for a tangible research material...MTAs.  Addgene does a lot to ensure that we can facilitate a fast and agile MTA process. This is important...deposits in our repository. From 2004 to 2023, we facilitated 784,795 MTAs with 8,798 organizations in 111 ...111 countries. From 2019 to 2023, we averaged facilitating 89,477 MTAs per year.  The MTA Process at Addgene...MTAs for plasmids in our repository. Instead, we facilitate the signing of MTAs by the Provider and Recipient...signature on any subsequent MTA for that plasmid facilitated by Addgene — basically, instead of having to ...shared reagents should be used. Setting clear expectations from the beginning makes the sharing process...

...others.  Meditate. Meditating during the day is a great way to take a mental break, while meditating right...Organize your data and prepare publications or your dissertation Organize data that you already collected into...into figures for papers or presentations. Learn a new skill for analyzing and visualizing your data, such...needing to have collected data. Work on your dissertation, even if your defense isn’t for a while. Getting...Data Hub to help the research community. Set expectations for your productivity Set a routine and stick...

... cutting edge genomic technologies to map the mutations that are present, and use prior knowledge (data...One central bottleneck is that most recurrently mutated cancer genes are rare and most of the individual...characterized? Some have developed exciting computational algorithms, such as SIFT or PolyPhen to approach...well as reference cDNA clones mapping to well-annotated cellular pathways). It wasn’t clear whether gene-agnostic...High-throughput Phenotyping of Lung Cancer Somatic Mutations. Berger AH, Brooks AN, Wu X, et al. Cancer Cell...perform additional functional assays for these mutations and map other cellular functions and phenotypes...to build saturating libraries of all possible mutations in many major cancer genes. These should be used...

...track of computation work comes with new sets of challenges. One challenge with computational analyses... of biologists are finding themselves using computational approaches to analyze large data sets (myself...what works, and what does not work. Careful documentation will keep you on task and will prevent you from...especially since you’ll want to dig in to the computation, but they will serve you well as you perform ... the analyses: Goal: Before you begin, briefly state the goal of the analysis, including background on... you to perform this analysis. Having a clearly stated, specific goal for each analysis will help you ...

... homologous recombination to facilitate simple gene replacement/mutation. Yeast expression plasmids used...microorganisms, yeast have both a stable haploid and diploid state which is useful for genetic analysis, as well as...apparently independent of a known specific sequence) dictate the replication of these vectors. S. pombe plasmids...missing nutrient. Scientists can exploit these host mutations by including a copy of a functional gene which...This can complement ura4- S. pombe, but the complementation is weak. yes   LYS2 L-lysine yes - Grow on... S. pombe  no no *FOA in fission yeast induces mutation in the ura5+ gene in addition to ura4+. leu1...

... DIO (Double-floxed Inverse Orientation) or DO (Double-floxed Orientation)) is a powerful tool that provides...channelrhodopsins that, in turn, trigger neural excitation, whereas yellow light activates halorhodopsins...resulting fusion gene is inserted in the antisense orientation with respect to the promoter to prevent its expression...recognition sites (e.g., loxP and lox511) in opposite orientations. Since Cre does not cause recombination between...therefore locking the opsin into the correct orientation to be transcribed (Figure 2) (Sharma and Zhu,... arousal in mice previously subjected to steady-state general anesthesia (Taylor et al., 2016). These ...strains of Halobacterium halobium differing in pigmentation." Biochemical and biophysical research communications...

...: Summary of Rosella’s excitation and emission spectra. Component Excitation Emission pH Range DsRed.T3... inhibitor. While this method does provide a quantitative measure of autophagy, it only measures bulk ...final read out of this assay. This is also a quantitative measure of autophagy, but, like the protein ...DsRed.T3. See Table 1 for a summary of Rosella’s excitation and emission spectra. DsRed is the permanent ... 508 nm ~6.5 - 9 *Optimal sequential excitations wavelengths as determined by Rosado et al: see... T., & Klionsky, D. J. (2008). Chapter 3 The Quantitative Pho8Δ60 Assay of Nonspecific Autophagy. Methods...

...and methylated states. Thus, this system is a reliable tool in reporting methylation status of these sites...the headlines, with sotires like the potential devastation of the palm oil industry through epigenetic effects...grandchildren; in this study,epigenetic changes were statistically significantly associated with reduced life-span...syndrome is characterized by low muscle tone, short stature, cognitive disabilities, problem behaviors, and...epigenetics; techniques include chromatin immunoprecipitation, fluorescent in-situ hybridization, methylation-sensitive...methyllysine binding domain causes a structural change (facilitated by the flexible linker) that brings the fluorescent...

...and immunology. They can be used for qualitative and quantitative measurement of protein expression in...protein via fluorescence or chemiluminescence, precipitate the protein out of solution, or isolate cells...epitope of the target protein. However, small mutations in the DNA of the hybridoma can occur, known as...types or treatment conditions. They can be used quantitatively if you also load the gel with appropriate controls...media from cultured cells. ELISA can be used to quantitatively measure the amount of protein present. The ...ELISA (Enzyme-linked Immunosorbent Assay). Immunoprecipitation Sometimes, instead of just detecting the ...immunofluorescence), or by a colorimetric reaction facilitated by an enzymatic reaction. Figure 5: Immunofluorescence...

...assays include immunoprecipitation mass spectrometry (IP/MS) and chromatin immunoprecipitation (ChIP). In ...In IP/MS, a target protein is first immunoprecipitated - along with its associated protein complexes - ...tag” such that these tagged proteins can be immunoprecipitated using a standard tag-specific antibody (Jarvik...double-strand breaks (DSBs). DSB repair by HDR facilitates in-frame insertion of tag-encoding sequences ...efficiency across different stem cell types. This has facilitated easy generation of clonal knock-in lines that... design, remember to incorporate PAM-blocking mutations (e.g. convert the NGG PAM to NGC in your repair...2018), however, if you have any questions, do not hesitate to contact me at [email protected]. Keep ...

...2xNLS) and c-terminal (Cas9(C)-2xNLS) fragments facilitates target DNA cleavage upon co-expression and spontaneous...has been cumbersome. To overcome many of the limitations intrinsic to DNA-guided systems such as TAL effectors...or invert a specific chunk of DNA. However, a limitation to this strategy is that each "bit" of information...This leads to 2048 (211) possible combinations of states. Each of the integrases is supplied on its own ...more powerful LID, the authors recently used computational protein design, phage display and high throughput...system. How does it work? RUSH is an ingenious two-state assay based on the reversible interaction between...synthase pair is encoded on a single plasmid and facilitates the incorporation of a variety of bio-orthogonal...

...but unsure how best to prepare. Maybe you’re a presentation veteran, but have difficulty answering questions...what you would like to say to help build your presentation. For example, when thinking about discussing... Practice, practice, practice: Rehearse your presentation with people inside and outside your field. Your...and PI can provide feedback on your technical presentation, but those outside of your field can point out...you a chance to determine how to tailor your presentation so that it can be understood by your audience...It’s likely you won’t know every answer – don’t hesitate to be honest and tell your audience just that....attendees can get sucked in to thirty-minute-long presentations, which constrains the number of posters you ...

...single protein, computationally described as a fusion of four Cas7 proteins and a putative Cas11-like protein...patient's specific mutations. With a drag and drop approach, however, the specific mutations wouldn’t matter...evolution, were also working on the project. Using computational observations, Koonin and Makarova found that...which can be caused by thousands of different mutations, all found in the same gene. If we only have small...matter – the mutated copy could be replaced by simply dragging and dropping in a functional copy of the entire...

...the late 1980s, scientists even created a codon adaptation index (CAI) that is based on the codon usage ...codon bias. The idea is that scientists can make mutations throughout a gene of interest based on an organisms...) to name a few. This could affect synonymous mutations which have even been linked to some diseases. ...human genes contain a region where a synonymous mutation can be harmful (Sauna and Kimchi-Sarfaty, 2011...tabulated from international DNA sequence databases: status for the year 2000. Nucleic Acids Research 28:292... Understanding the contribution of synonymous mutations to human disease. Nat Rev Genet 12:683–691. https...10.1038/nrg3051 Sharp PM, Li W-H (1987) The codon adaptation index-a measure of directional synonymous codon...

...template to create mutations HDR donors can contain insertable sequences or specific mutations, along with homologous...sequence at the DSB, or it can introduce specific mutations into the damaged DNA if these modifications exist...HDR efficiency for more details on how "Cut-to-Mutation distance" affects editing efficiency. One important... cutting and repairing your target DNA until a mutation is introduced disrupting one or both of these ...sequences. To prevent re-cutting and the resulting mutations after an HDR event, ensure the gRNA or PAM site...off-target effects. A technique that pushes the size limitations of ssODNs is Easi-CRISPR (Quadros et al., 2017...

... introduce naturally occurring cancer-causing mutations into the endogenous allele of the STAG2 tumor ... introducing non-sequence specific frameshift mutations through aberrant double strand break repair, it...knockins, it is not always ideal. Because of these limitations, the AAV approach is best suited for tissue culture...epitope tags, or the introduction of dominant mutations of a gene into the endogenous allele in cultured...template to mouse zygotes and thereby correct a mutation that causes Hemophilia B. Unfortunately, work ...applications that require large gene fusions. The current state-of-the-art in combining the AAV and CRISPR-based...segregation in human cells harboring tumor-derived mutations in STAG2." PLoS genetics 12.2 (2016): e1005865...

...biotin ligase, BirA*, with a site mutation (R11G). This site mutation destabilizes the ligase, allowing...the E. coli version. It also contains a single mutation (R40G) in a conserved residue in the biotin catalytic...ascorbate peroxidase for proximity labeling A major limitation of APEX was its low sensitivity. Sensitivity ... two mutants, the 35 kD TurboID, which has 15 mutations compared to BirA and the 28kD miniTurbo, which...which has the N-terminus domain deleted and 13 mutations compared to wild type BirA. TurboID and miniTurbo...interactions Split-BioID combines a protein fragment complementation (PCA) technique with proximity labelling to...

...fluorescence that is generated allows for qualitative and quantitative measurement of processes including RNA...photobleaches, DFHBI can only be used for qualitative, but not quantitative, studies. Additionally, Spinach, the... In the search for something more stable for quantitative imaging, Ming Hammond’s group created Spinach2...photostable than Spinach and Broccoli, and useful for quantitative studies. Specifically, Corn was developed to... and both TO1-biotin and TO3-biotin contain an acetate modification that reduces nonspecific binding with...

...within this kit contain a variety of promoters to facilitate gene expression in bacteria, yeast, mammalian...His, Avi, and GFP) as well as protease sites to facilitate protein purification. We hope this kit will allow...important microalgae: intrinsic suicide switch implementation in cyanobacterium Synechocystis sp. PCC 6803...Furthermore, some plasmids contain peptides to facilitate protein folding and/or protease sites to remove...The gene set was carefully chosen to perform statistically relevant benchmarking of cell-free expression...By counting DNA sequence reads, FREQ-Seq can quantitatively determine allele frequencies across timepoints...selective pressures: Miller, Corwin, et al. "Adaptation of Enterococcus faecalis to daptomycin reveals...

...purified GFP could be imaged in the green state after excitation at 488 nm, photoconverted with 405 nm light... light, and then captured in the red state after excitation at 561 nm. This imaging process worked well...S65T GFP, and no red color was observed after excitation at 488 nm, indicating that spontaneous conversion...al., Sci Rep. 2015 CC BY 4.0 Applications and limitations of GFP photoconversion One concern with photoconverting...

...knock-ins of patient mutations which are variants of unknown significance – mutations with unclear effects...with defined, straightforward causes (a specific mutation in a known gene = disease onset) may be best served...accessible, fast, and easy. In cases where the mutational basis of a disease is known – repeat expansion... can aid in generation of multiple edits or implementation of expression systems to accurately mimic a...CRISPR could even be used to generate patient mutations and then test both individual and combinatorial...

...detect proteins in different states. For example, in immunoprecipitation (IP) the antibody interacts with... for protein interaction studies such as immunoprecipitation and pull-down assays.  Westerns are divided... with a protein in its native state while a western blot detects denatured polypeptide chains. The protein...epitopes, differ between a protein in its native state versus the polypeptide chain. Consequently, antibodies... way to gauge protein abundance they provide qualitative measurements. If you require precise quantification...determine protein interactions following immunoprecipitation and are a common readout for subcellular ...

... H161Y, P163T, I175Q, C176T and N177F mutations). Its excitation maximum is reported as 603 nm, while ...aptamers and MS2-binding fluorescent proteins were the state of the art in RNA imaging in vivo. These new “Riboglow...et al. PLoS One, 2016) and tested a series of mutations. Through this process mCarmine was developed (...several cell types is the lack of a rapid and quantitative editing assay. To help accelerate the expansion...-UGI-mediated activity edits out a frame-shift mutation within the mCherry reporter. Quantification of...

... job asks for experience in “statistical analysis”, mention “statistical analysis” (word by word) in your...general format of a resume and the different presentation styles. Wikipedia does a good job summarizing...handled large data sets and are familiar with statistical analysis. If you have done fluorescent imaging...publishing or patent application processes? How about facilitating a collaboration between research groups? Perhaps...universities make their job postings and salary expectations public, so start by looking at these university...

... that cluttered our old plasmid maps. Clearly Annotated Features, Enzymes, and Primers - The new maps ...SnapGene’s software positions restriction enzyme annotations throughout the map at locations better aligned...map, you also have the ability to to download the static image file (PNG, see above) for your own reference...sequencing results: On this page you can download an annotated GenBank File or a SnapGene file for the sequences...of choice. If the depositor provided a custom annotated GenBank file, this can be found on the main plasmid...hosted here retain all the features shown in the static maps discussed above, but also have interactive...

...incorporation of a desired mutation into your plasmid of interest by PCR generates mutated plasmids with no methylation...only the non-mutated and methylated template is destroyed leaving behind a pool of mutated plasmids which...dam-/dcm- strains may have an increased rate of mutation (as these would also be deficient in mis-match...Your Sequencing Results Use REPLACR Mutagenesis to Mutate Your Plasmids Resources on Addgene.org Check ...

...the immune mechanisms of the host. The cause, manifestation, diagnosis, recurrence, and treatment of cancer...molecular targets under research include TIM-3, LAG-3, STAT-3, EGFR, CD20 and CD73 on specific solid as well..., tumor penetration, inadequate concentration, mutation of cell surface targets by tumor cells, are a ...currently the only FDA-approved DC therapy for prostate cancer and is aimed at increasing life expectancy...radiotherapies have caused increased death receptor presentation on tumor cells thus making these cells more ...differ in practically every facet starting from manifestation to their specific interactions with different...the most important: How will cancers evolve and mutate based on the immunotherapeutics we use on them?...

...cartoon of the protein.) In the middle of my presentation, lost in a sea of data, my mind went completely...taught me an incredibly valuable lesson: Your presentation will be better if you take time when you need...need it, rather than trying to push through a state of panic. Now, if I feel myself getting too nervous...sense. As tempting as option two was in my panicked state, only option one would let me accomplish my goal...often as needed until you get to the end of the presentation, at which point you will have achieved your ...gig. Being able to give a clear, informative presentation, speak at conferences, or even just provide ...

...light-sensitive protein QuasAr2 and systematically mutated it. Using a robotic picking approach to select ...2015) or ratiometric Bimolecular Fluorescence Complementation (rBiFC, Grefen and Blatt, 2012). The 2in1 Plasmid...vectors with all possible combinations of tag orientation (NN, NC, CN, and CC). The 4 kits also contain...proteins that bind to either the light- and dark- states of the photoswitchable cPYP, which changes in conformation... partners that bind to only the light- or dark- state of the photoswitchable proteins offers a powerful...binders (which bind to either the light- or dark- state of the photoswitchable proteins) fused to mVenus...

...dominant negative mutation in MLH1 (PE4 = PE2+MLH1dn and PE5 = PE3+MLH1dn). Silent mutations added near the...the Cas12f scaffold gRNA and making strategic mutations in the DNA binding cavity of Cas12f. The CasMINI...  and base editing, whereas a different set of mutations (the “V3.1” version) works best for genome editing...protein PEG10 specifically binds its own mRNA to facilitate its packaging into virus-like particles. Feng...boosts the efficiency of prime editing to correct mutations in a variety of target genes and cell types. Their...