...various components of the immune system. Figure 2: In-vitro models for immunogenicity prediction of ...drug reaction. This system would somewhat act like a 2-step verification. All approaches have their own limitations...Future Enhancements. Frontiers in drug discovery, 2, 952326. https://doi.org/10.3389/fddsv.2022.952326Joubert...ADAs can inactivate the drug and increase drug clearance, lowering its efficacy and increasing the risk...Efforts to predict T cell epitopes focus on characterizing those epitopes as CD8 or CD4 epitopes, as well...

...and a source of sdAbs (Figure 2) (Xu, et al., 2021).      Figure 2: Genetically modified mice offer... from camelid mice and llamas neutralize SARS-CoV-2 variants. Nature 595:278–282. https://doi.org/10.1038...2024 the International Year of Camelids. The declaration is intended to raise awareness of the economic...

... Opto-Cre-Vvd is fast and can cut in as little as 2 hours.  Sheets, MB, et al., ACS Synth Biol. 2020....they investigated three Casɸ orthologs, Casɸ-1, Casɸ-2, and Casɸ-3. This system is notable for several reasons...a minimal PAM requirement. For instance, for Casɸ-2 the PAM sequence is 5’-TBN-3’ (where B is G, T, or...deposited by Leopold Parts. This library delivers 2 randomly paired guides per construct and allows a ...crRNA-guided DNA cutting, unlike other well-characterized Cas enzymes, which rely on multiple active sites...

...developed around how they replicate (Figure 2). Figure 2: Baltimore replication classes. Image credit...safer viral vectors that can be used in BSL-1 or BSL-2 labs. Viral vectors are viruses that have been genetically...genes for the required proteins are packaged in separate plasmids and expressed via cells cultured with...

...calcium biosensor, so that the (1) binding of calcium, (2) allosteric rearrangements of the sensor, and (3) ...user-controlled illumination with violet light and (2) exposure to increased intracellular calcium. It has...dynamics in vivo. Neuroscience Research. 2021 Aug 1;169:2-8. Suzuki J, Kanemaru K, Iino M. Genetically encoded...experiment with a sensor that is tuned to your parameters.   Kinetics How fast is the process you would...used calcium reporters with different kinetic parameters. There are three GCaMP types based on their kinetics...available with improved kinetic and sensitivity parameters. Sensitivity and Localization The calcium concentrations...to go. Pro tip: Vectors with ready-to-use AAV preparations are labeled with the green virus particle on...

...CombiGEM lentiviral vector backbone pAWp12 (Figure 2). Cloning from the pAWp28 storage vector to the pAWp12... before beginning your experiments. Figure 2: Strategy for assembling barcoded combinatorial gRNA...PMID: 25114216. PubMed Central PMCID: PMC4151723. 2. Doench, John G., et al. "Optimized sgRNA design to...has been successfully applied to functionally characterize combinatorial gene knockouts generated via multiplexed...antibiotic-resistant bacteria enabled by massively parallel combinatorial genetics." Proceedings of the National...PMC3972032. 10. Wong, Alan SL, et al. "Massively parallel high-order combinatorial genetics in human cells...

... was first discovered and characterized in an unrelated worm (C. elegans) [2]. Delivering dsRNA matching...neoblast cells and the gene Prohormone convertase 2 (pc2) labels the central nervous system. Evaluating...223). Link: http://abt.ucpress.edu/content/79/3/208 2. Fire, S. Xu, M.K. Montgomery, S.A. Kostas, S.E. Driver...Ross, Sofia M. C. Robb, Sarah A. Elliott, Kimberly C. Tu, Alejandro Sánchez Alvarado, The American Biology...riboprobes are created, the WISH protocol involves preparation of the tissue sample, several washes, and temperature...PMC304099. Additional Resources on the Addgene Blog Arabidopsis in Education Learning Synthetic Biology with ...

...point could be a molar ratio of 1:2 (plasmid:insert), but this parameter has to be optimized according to... working with is correct. It's also best to use 1-2 ug of the vector for digestion. PCR Design primers...colonies (5-10) and grow them in a small culture volume (2 or 3 ml) containing the corresponding antibiotic, ...to be strategy specific, there are few general parameters that you can adjust: temperature and time of ...

...required to make a plasmid. In 2023 (and at closer to 2 million plasmids shared than not!), we think it’s ...time does it take to make a plasmid? Figure 2: Survey results from a Twitter poll run in 2023, n...go wrong, replace them for free. Plasmids ship out 2-3 days after ordering, making it a toss-up as to whether...relevant work in your field! However, there’s no guarantee you’ll get a response on the first try (or any...at a time, try constructing several plasmids in parallel. That way, you can run one PCR, one gel, etc. ...

...or non-homologous end joining (see Figure 2).  Figure 2: Schematic representation of integration...due to insertional mutagenesis (Howe et al., 2008). 2. Transient expression of transgenes/shRNAs If you ...vectors: progress and applications. Gene Therapy, 17(2), 150–157. https://doi.org/10.1038/gt.2009.135. PubMed...efficiently during infection. Journal of Virology, 70(2), 721–728. https://doi.org/10.1128/JVI.70.2.721-728.1996...our knowledge here, the Pol gene encodes three separate enzymes that are proteolytically processed from...

... used. Follow the manufacturer’s instructions.   2. Isolate Your Insert and Vector by Gel Purification...DNA concentration alone. One method is to conduct 2 ligations for each plasmid you are trying to create...containing the ligation mixture with backbone alone; 2. Transformation containing ligation mixture with insert...colonies than the vector + insert plate (unsuccessful 2), or roughly equivalent numbers of colonies on each... or equivalent to backbone + insert, unsuccessful 2 and 3 above), you can try ligating the recipient plasmid...slower than smaller linearized fragments. You can separate your backbone away from any inserts cut out of...

...of other Cas proteins found in the so-called class 2 CRISPR-Cas systems. There are 24 variants of the cas14...the presence of infectious organisms (ex: SARS-CoV-2) and genetic mutations. By providing a gRNA specific...Department of Energy’s Joint Genome Institute for uncharacterized genes that were proximal to a CRISPR array ...presence of a predicted RuvC nuclease domain, characteristic of CRISPR-Cas enzymes. In contrast to other...

...research experiences to undergraduate students (1, 2), but educators face significant challenges in designing... by western blotting in other experiments (Figure 2).   Complementation reveals conservation of function...Biology Education: A Call to Action, Washington, DC. 2. President's Council of Advisors on Science and Technology...Functional characterization of the Saccharomyces cerevisiae genome by gene deletion and parallel analysis...cerevisiae (Figure 1) occurs through a well-characterized pathway with multiple enzymatic steps (5). In.... cerevisiae met deficiencies (manuscript in preparation), indicating that Met protein functions have ...

...AAVs in the clinic Thus far, AAV vectors AAV1, AAV-2, AAV1-AAV2 hybrids, AAV-6, AAV-7, AAV-8, AAV-9 and...neutralizing antibodies against some AAV types, AAV1, 2, 3 and 5 are thought to be as high as 70% among the... 10.24 (2003): 1999-2004. PubMed PMID: 14566358.  2. Hirsch, M. L., et al. "Self-complementary AAV mediates...common cytokine receptor gamma chain (c) and is characterized by a lack of immune cell development and function...first discovered as a contaminant of adenovirus preparations. It is an icosahedral, non-enveloped virus that...repair via homology directed repair (HDR). This characteristic makes AAV vectors especially amenable to correcting...caused by mutations in the gene RPE65. It is characterized by childhood onset of blindness and was thought...

...possible when each of the XFP genes is introduced at ~2 copies per cell per color, following a Poisson distribution...(prepared with Addgene #104110 - #104112) (Figure 2). We’ve also created constructs with containing chemical...AAV titer - The XFP genes have to be introduced at ~2 copies per cell per color to achieve the highest color... levels of XFPs - Reduce the amount of tTA. Paradoxically, too much tTA leads to a reduced expression ...

...human transcription factors such as PAX7, PAX6, AP-2 alpha and ISLET-1. In addition, we distribute monoclonal... example through the use of less toxic adjuvants; 2) generating complex antibody chips that allow assessment...variable, allowing exquisite specificity. This characteristic allows monoclonal antibodies to be used as ...

...the mixed sample in our pipeline (see Figure 2, sample 2). When we blindly analyzed AAV-68544, we were...look for potential contaminants. The analysis is a 2-step process using Geneious software: First, the individual...Léger A (2015) Advanced Characterization of DNA Molecules in rAAV Vector Preparations by Single-stranded Virus...especially important when producing multiple vectors in parallel because the risk of accidental errors increases... be used to identify DNA contaminants in AAV preparations and provide information on these DNA species...pioneering the systematic use of this new QC assay to guarantee that you receive the best AAV vectors for your...Identity of Recombinant Adeno-Associated Viral Preparations from Viral DNA Extracts. Human Gene Therapy ...

...Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system." Cell (2015). PubMed PMID: 26422227... Find plasmids from this publication at Addgene. 2. Kleinstiver, Benjamin P., et al. "Genome-wide specificities...Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell. 2015 Sep 23. pii: S0092-8674...also called Cas12a. In 2015, Feng Zhang’s lab characterized two Cpf1 nucleases, distant cousins of well-...Until recently, two major elements of Cpf1 characterization were missing: on-target and off-target editing...efficiency. Kim et al. and Kleinstiver et al. characterized genome-wide editing efficiency of two Cpf1 orthologs...lower than that of the widely used SpCas9 and comparable to SaCas9. As seen with Cas9 orthologs, Cpf1 ...

... fluorophore excitation by absorption of a photon 2. Energy transfer from the excited donor directly to...PMID: 24194740. PubMed Central PMCID: PMC3810607. 2. Bajar, Bryce T., et al. "A Guide to Fluorescent Protein...PubMed PMID: 10500161. PubMed Central PMCID: PMC18018. 2. Miyawaki, Atsushi, et al. "Fluorescent indicators...various papers to make the decision easier. 1. Müller, Sara M., et al. "Quantification of Förster resonance ...donor and acceptor fluorophores are fused to two separate proteins. (In contrast, intramolecular FRET probes...error in suboptimal or undetectable FRET, well-characterized FRET reference standards can be used to validate...

...therapy had fallen - plummeted really -  from grace 2 years prior with the death of Jesse Gelsinger in an... the whole field to a grinding stop and only now, 2 decades later, is it finally back in the limelight...the other hand, have broad tropism in the eye. AAV-2, the first identified serotype and the one used in...reading 1. FDA approves gene therapy for blindness 2. Gene Therapy emerges from disgrace to be the next...occurring animal model, the RPE65−/− dog, was characterized and found to suffer from early and severe visual...

... However, if you have a larger knock-in (200 bp – 2 kb), a dsDNA donor will need to be used due to synthesis...accidentally change your protein of interest.   Fig. 2: Stop re-cutting with PAM and guide RNA disrupting... the other repair mechanisms Mammalian cells have 2 primary repair mechanisms besides HR - Non-homologous.... Com., 8(2024), (2017). 10.1038/s41467-017-01836-2 Charpentier, M., Khedher, A. H. Y., Menerot, S., Brion...recognition sequence in the donor DNA, you can guarantee there won’t be re-targeting after HDR. If your...

...indicated they wanted to see more negative data (Figure 2.) At the same time, when scientists who support open...is not only housed, but encouraged.    Fig. 2: Results from a survey conducted by Addgene in 2021...detailed methods and image descriptions, a minimum n = 2, and well labeled high- and low-magnification images...gene therapy trials. Human Gene Therapy Methods, 26(2), 71–76. https://doi.org/10.1089/hgtb.2015.040 Initiative... in Adeno-Associated Virus Serotype 9 (AAV9) Preparations. Human Gene Therapy Methods, 30(6), 214–225....

...procedure to be performed using a simple two-step, 2 hr protocol prior to transformation (Figure 1). As...Primer design Using specific primer design (Figure 2), IVA cloning can be used to perform any plasmid modification... for each type of modification is shown in Figure 2, and is discussed in more detail below. The main requirement...Biotechniques 10.1 (1991): 62-66. PubMed PMID: 2003926. 2. Bubeck, Peter, Monica Winkler, and Wilfried Bautsch...another simply amplify the vector and insert with separate primer pairs in the same PCR. Add homologous sequences...

...create the 2nd-generation lentiviral system (Figure 2). The HIV genes that do remain are very important ...1996 Apr 12;272(5259):263-7. PubMed PMID: 8602510. 2. A third-generation lentivirus vector with a conditional...CH, Humeau LM, Rebello T. J Gene Med. 2013 Feb;15(2):78-82. PubMed PMID: 23322669. 6. Genetic treatment... Banasik MB, McCray PB Jr. Gene Ther. 2010 Feb;17(2):150-7. PubMed PMID: 19847206. Additional Resources...Mulder GJ, Polack A, Bank A, Soulier J, Larghero J, Kabbara N, Dalle B, Gourmel B, Socie G, Chrétien S, Cartier...

... to DFHBI in the presence of that metabolite (Fig.2). Spinach eventually evolved into Spinach2, a more...PMID: 25713381. PubMed Central PMCID: PMC4364232. 2. Baker, Monya. “RNA imaging in situ”. Nature Methods...emerging environmental pollutants.” Front Chem. 2014; 2: 41. PubMed PMID: 25019067. PubMed Central PMCID: ...biosensors.” TrAC Trends in Analytical Chemistry 27(2):108-117 · February 2008. 11. Svensen, Nina and Jaffrey...would bring the relevant ends together rather than separate them (Fig. 1b). One can also make “signal-off”...

...droplet in each well (Figure 2). A clean ddPCR should have a clear separation between positive (blue) and... to 2 minutes and the denaturation time to 1 minute has been shown to increase droplet separation (Witte... qPCR is that the results can vary by a factor of 2. This means that if you set up two identical assays...getting a clean separation between positive and negative droplets with these parameters, here are a few...Since Addgene began producing AAV vector preparations, we have used quantitative PCR to evaluate the ... for ddPCR set-up. Prepare the master mix in a separate area from where you prepare your sample dilutions...your PCR For a good starting place on what PCR parameters to use, see Lock’s seminal paper on AAV titration...

...3 plates at different densities for selection). A 2% agarose solution is then melted (and thereby sterilized...96-well assay plate. We generally freeze at least 2 ampules each of 4 clones with different levels of ...of agarose, which is then overlaid with the firmer 2% agarose gel solution discussed above for picking....concentrations of salts and glucose. If we circled well-separated clones containing more than 50 cells (visible ... should contain at least 50 cells and be well-separated. If the growth rate/transfection efficiency is... with this!) Make sure your colonies are well separated. For this, you need to have some idea of the rate...individual cell clones with different expression characteristics from a population of transfected clones. The...

...phosphorylation events in the proteomes of higher mammals (2). How can we begin to understand the importance of... the open reading frame (ORF) of interest (Figure 2). 1. The phosphoserine OTS The backbone of the phosphoserine...optimized Sep-OTSλ and can recommend its use (6).  2. C321, a specialized bacterial host strain for pSer-incorporation...biology4.5 (2002): E127-E130. PubMed PMID: 11988757. 2. Hornbeck, Peter V., et al. "PhosphoSitePlus, 2014...phosphorylated intermediate of a protein for characterization. However, phosphoproteins are often low abundance...strategies; expression should be optimized for parameters such as induction temperature and length of induction...

...mammal called alpaca could neutralize the SARS-CoV-2 virus. But while the early results were promising,...receiving dozens of requests every day.    Figure 2: When a researcher needs a collaborator for a specific...Crowdfight will: 1) Find an expert with the right skill; 2) Confirm the expert’s suitability and availability...diseases, so we were left with no choice but to quarantine in our houses, feeling helpless about the situation...

...to (1) run a DNA gel to confirm fragment size and (2)  incubate with RNAse and Proteinase K to purify the...proteins of interest to any DNA they were bound to (2) fragmented the DNA (3) used antibodies to specifically...adjustments, to get everything just right!  Fig. 2: Goldilocks is actually celebrating a successful ChIP...remain bound to each other until you are ready to separate them. Most proteins are not likely to remain attached...

... Jan;20(1):81-89. PubMed. 2. Bauer DE, Kamran SC, Lessard S, Xu J, Fujiwara Y, Lin C, et al. An erythroid...with TALENs, in collaboration with the Porteus lab [2]. With the advent of CRISPR/Cas9, we began to explore... inferred based on correlative biochemical characteristics and ectopic reporter assays. We have been interested..., identifying these deletions, and finally characterization. Read and download the full protocol from ...Dass A, Yien YY, Chung J, Masuda T, et al. Characterization of genomic deletion efficiency mediated by...

...the result in Figure 1 could look like this: Figure 2: Expression of Plasmid B (as internal control) This...down expression of Gene X but, as shown in Figure 2, this was likely due to the original transfection ...PMID: 20074371. PubMed Central PMCID: PMC2817684. 2. Replicates and repeats—what is the difference and...multiple separate wells (within the same plate) with purified plasmid from the same aliquot/preparation. The...co-transfected with the test plasmid or transfected into a separate well of your cells. Regardless, the amount of ...should be consistent over time and from different preparations of the purified test plasmid expressing the ...Using our example above, we would test different preparations of Plasmid A in different aliquots of cells ...

... is also gaining a lot of new traction lately (1, 2), depends on knowledge of the structure(s) a given...per plate) across 20 fresh selective plates. Step 2: Express random phage DNA fragments in E. coli Now...microbiology 23.4 (2015): 185-191. PubMed PMID: 25708933. 2. Cooper, Callum J., Mohammadali Khan Mirzaei, and ... biology 34 (2015): 69-77. PubMed PMID: 26275959. 2. Weigele, Peter, and Elisabeth A. Raleigh. "Biosynthesis... to be explored and exploited. With better characterization, these proteins could function as surrogate...surrogate antibodies for identifying and characterizing sugars and sugar-containing molecules in molecular biology...colonies of interest, sequence the inserted DNA and characterize your new putative host cell-binding proteins...

.../10.1038/s41587-024-02268-2 Schep, R., Brinkman, E. K., Leemans, C., Vergara, X., van der Weide, R. H.... flanking sequences to their input string (Figure 2). This string can then be used as input for the prediction...top 5) and test them in their labs. Figure 2: Guidelines to predict pegRNA efficiencies with pridict.it...we can evaluate tens of thousands of designs in parallel. To do this, we performed high-throughput ‘self-targeting...the TRIP (Thousands of Reporters Integrated in Parallel; Akhtar et al., 2013; Schep et al., 2021) method...Assayed by Thousands of Reporters Integrated in Parallel. Cell, 154(4), 914–927. https://doi.org/10.1016...., Benvenuto, E., Schmidheini, L., Schep, R., Damodharan, T., Balázs, Z., Janjuha, S., Ioannidi, E. I....

...upstream of an opsin gene (e.g., channelrhodopsin-2, ChR2) fused to a reporter gene (e.g., mCherry) to...the correct orientation to be transcribed (Figure 2) (Sharma and Zhu, 2014). How does FLEx enable strong... Abdallah, Khaled, et al. "Adeno-associated virus 2/9 delivery of Cre recombinase in mouse primary afferents...Deniz, et al. "A FLEX switch targets Channelrhodopsin-2 to multiple cell types for imaging and long-range ...vectors to answer this question, mapping and characterizing the activity of the DA neurons of the VTA. ...brain area, demonstrating that it is possible to separate neuronal inputs to induce aversion- or reward-...PMC4971574. Hooper, Andrew, and Jamie Maguire. "Characterization of a novel subtype of hippocampal interneurons...

...) 38 kb Cells HEK-293T Sf9 Time to collection 2–4 days 5–10 days Purification Ultracentrifugation...Centrifugation Titration time ~1 day (AAV); ~1 week (lenti) 2–10 days Initial titers Higher Lower Resources... a factory to produce adeno-associated virus type 2 vectors. Human Gene Therapy, 13(16), 1935–43. https...transfection). This differs from typical insect cell preparations, which we talk about below. Processing The viral...Plasmids and Resources References Shaw, G., Morse, S., Ararat, M., & Graham, F. L. (2002). Preferential transformation...

...experiment using smFPs and standard neuronal tracers. 2. Visualization of neuronal sub-cellular structures...similar proteins. For example, N-cadherin (cadherin-2) is a postsynaptic cell adhesion protein that plays...PMID: 25915120. PubMed Central PMCID: PMC4573404. 2. Shepherd, G.M. (2004). In "The synaptic organization...STORM imaging, and electron microscopy. Sample preparation for these techniques sometimes makes it more ...retain their ability to be detected after sample preparation better than standard proteins tags like GFP. ...smFPs in the future. References 1. Viswanathan, Sarada, et al. "High-performance probes for light and ...

...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...inherent brightness and spectral characteristics of the moxFPs are comparable to the non-optimized parental...protein activity. We encourage investigators to characterize FP-fusion proteins functionally relative to ...

...revert back to a pluripotent stage (iPSC formation) [2, 3]. The iPSCs then proliferate and redifferentiate... more common delivery methods include retrovirus [2], lentivirus [4], adenovirus [5], Sendai virus [6]...2006. 441(7097): p. 1061-7. PubMed PMID: 16810240. 2. Takahashi, K., et al., Induction of pluripotent stem... Pluripotent Stem Cells. Cell Stem Cell, 2015. 17(2): p. 178-94. PubMed PMID: 26189426. 15. Si-Tayeb, ...induced pluripotent stem cells. Stem Cell Rev, 2015. 11(2): p. 242-53. PubMed PMID: 25578634. PubMed Central... to regeneration. Nat Rev Mol Cell Biol, 2011. 12(2): p. 79-89. PubMed PMID: 21252997. 37. Merrell, A....adapted from Zhou and Melton, 2008, [43]): Characteristic Reprogramming+ Transdifferentiation in vivo...

...indicating primer dimers. (c) A double peak indicating 2 amplicons. Image credits (respectively): Zuzanna K...details for reproducibility’s sake!   Fig. 2: Setting up triplicates in multiple configurations...used method is the Standard Curve method. See Table 2, (adapted from Wong, 2018) for a comparison of different...Statistically defined log-linear phase   Table 2: A comparison of different qPCR analysis methods. ... for RNA quantitation. Biotechniques. 2000 Aug;29(2):332-7. doi: 10.2144/00292rv02. PMID: 10948434.Taylor...(Taylor, 2017). With the right conditions and parameters, some people find it can detect even smaller ...run a melt curve to validate your primers. This separate thermocycle allows you to measure the intensity...

... environmental sustainability in mind (Table 2). Table 2: Sustainability features highlighted in plasmid... and then clean-up the plasmid DNA (Figure 2). For most kits, the steps for lysing cells and removing ...together — no mixing and matching!   Figure 2: Plasmid isolation workflows are similar across kits...Sold Separately Yes, buffers and columns sold separately Some, waste tubes sold separately No...carry-over into the eluate, while other kits are guaranteed to yield endotoxin-free DNA. Once you know the...stages: lyse the bacteria harboring your plasmid; separate plasmid DNA from unwanted cellular “debris”; and...kits. Lyse your bacteria to release the plasmid, separate the cell debris from the plasmid, and then clean-up...

...under phase contrast starting day 2 after transfection.     Fig: 2: Expressing mARGAna in MDA-MB-231...engineering a cell line, cotransfect the mARG cassettes 2:1 (A:N-V) with PiggyBac transposase and outgrow the...for longer. Cultures induced with arabinose and colonies on arabinose-containing plates will be visibly... inducible expression systems in E. coli. The arabinose-inducible promoter pBAD and a low-copy number ...wide range of inducer concentrations (0.1-1% L-arabinose) without signs of toxicity or burden relative ...or on LB-agar plates containing glucose and L-arabinose at 37oC for at least 12 hours, or at lower temperatures...after the E. coli have colonized tumors, inject L-arabinose intraperitoneally and wait 24 hours before imaging...

...CcdB.” Biotechniques. 1996 Aug;21(2):320-3. PubMed PMID: 8862819. 2. Bahassi, EM., et al. “F plasmid CcdB... desired insert should not grow.   Figure 2: Image from http://teachline.ls.huji.ac.il/72682/tutorials...advantages that will be discussed in detail in a separate post. Focusing on the ccdB aspect, Gateway takes...

...discussed them in meetings and 2) creating a pictorial guide for our engineers (Fig. 2). Creating this guide was...communication has helped Addgene move from completing 2-3 large projects a year, on average, to completing...examples did not happen in the lab, they have parallels to collaborative lab work. Corralling context...

...evolutionary tree at least 6-7 times and on at least 2 separate continents. Mimulus (Andrea Sweigart) Monkeyflowers...presented in a series of quick (10 minute) or very quick (2 minute) talks so we got to hear from everyone. Addgene...Organisms, Part 1 Five Popular Model Organisms, Part 2 .... Sticklebacks demonstrate unusual parallel adaptation to disparate environments so can be used study ...difficult). Cuscuta (James Westwood) Dodders are parasitic plants that steal all their food from other plants...have economic importance as harmful weeds. The parasitic fusion is such that the two plants share RNA, ...

...A-B+ cells. If you look at the dot plot in Figure 2, you might assume that all cell populations are positive... the different populations. In the case of Figure 2, one population is B- and another two are B+. Our ...are in the lower right corner.      Figure 2: This flow plot has two populations that are B+ and...actual flow cytometry run, all markers are read in parallel.  Let’s imagine we’re looking for cells in a sample...markers to most specific, the first step is to separate cells from debris and roughly group the cells ...gated population, two types of FSC are used to separate single cells from doublets, ignoring anything ...population of B cells and dendritic cells is further separated into three different populations using conjugated...

...partitioning each type of plasmid into a separate daughter cells (Figure 2) (Ebersbach et al., 2005). However...to plasmid loss (Diaz et al., 2015).  Figure 2: The partitioning problem. For low copy plasmids...Plasmids. Microbiology and Molecular Biology Reviews 62(2): 434–464. Pubmed PMID: 9618448 Diaz R, Rech J, Bouet...model, termed the “handcuff” model, iterons of two separate plasmids bind to the same RepA molecules, forming...NTPase is recruited and “walks” each plasmid to a separate daughter cell. When plasmids are compatible, different...bind to each plasmid type, and different NTPases separate the plasmid pairs into the new daughter cells.... Pubmed PMID: 22153351 Shintani M, Sanchez ZK, Kimbara K (2015) Genomics of microbial plasmids: classification...

...-induced changes in root-to-shoot ratio in Arabidopsis, (2) expression patterns in plants with enhanced... go back and finish the review.  Peer-review step 2: Detailed assessment After this initial assessment... of Arabidopsis HapMap population in Amsterdam. Currently, she is a postdoc at KAUST, Saudi Arabia, in...Julkowska, a postdoctoral researcher at KAUST, Saudi Arabia. From the perspective of an author submitting ... specific repositories (Addgene, WormBase, Arabidopsis.info). These repositories will ensure that the ...guest blogger Magdalena Julkowska from KAUST, Saudi Arabia. Magdalena Julkowska did her Ph.D. on salt-stress...

...resistance to the herbicide chlorsulfuron (Homo line 2) or wild type (WT), two weeks after herbicide was ... CLIPf tag and a C-terminal SNAPf tag; see Figure 2), the authors developed a cost-effective expression... when associated with microtubules. Figure 2: CLIP-SNAP fusion proteins for single-molecule FRET... to drive prime editing in both tomatoes and Arabidopsis, suggesting the improvements may be broadly applicable...enables heritable desired edits in tomato and Arabidopsis. Nature Plants, 10(10), 1502–1513. doi: https...

...host range plasmids. FEMS Microbiology Letters, 348(2), 87–96. https://doi.org/10.1111/1574-6968.12241  ...mobilization of broad host-range IncQ plasmids. Plasmid, 62(2), 57–70. https://doi.org/10.1016/j.plasmid.2009.05.001...negative hosts. Biotechnology and Bioengineering, 106(2), 326–332. https://doi.org/10.1002/bit.22695  Schmidhauser...Understanding the characteristics of natural plasmids and how they function in nature can inform us on...trichosporium, Myxococcus xanthus, Neisseria spp., Paracoccous denitrificans, Pseudomonas spp., Rhizobium spp...Moraxella spp., Mycobacterium aurum, M. smegmatis, Paracoccus denitrificans, Pasteurella multocida, Porphyromonas...Escherichia coli, Gluconacetobacter xylinus, Paracoccous denitrificans, Pseudomonas fluorescens, P. putida...