...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...

...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...

...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...

...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...

...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...

...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...

...-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...

...improve solubility and stability (Figure 2).      Figure 2: Crystal structures of CRBNmidi alone...fluorescent tags in Caenorhabditis elegans. Genetics, 228(2), iyae126. https://doi.org/10.1093/genetics/iyae126...K., & Russell, W. C. (1987). Isolation and characterization of monoclonal antibodies to simian virus 5...matured considerably faster, with expression levels comparable to GFP during early embryogenesis (Figure 3)....Merritt D.M., Pe K., Cesar M., Hobert O. (2024) Comparative analysis of new mScarlet-based red fluorescent... and the LgBiT luciferase fragment (expressed separately) assemble and catalyze the luminescence reaction...

...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...

...Brainbow mouse neural labeling system. Brainbow-1 and -2 have been further refined to create Brainbow 3.2. ...M-CFP expression; these deletions are defined using 2 loxP variants (only identical loxP sites can mediate... each fluorophore. Third, Brainbow-1 and Brainbow-2 each contained a “default” state; for example, Brainbow...processes not previously visible with Brainbow-1 and -2. The general structure of Brainbow-1.0 is retained...fluorescent proteins to find those with ideal characteristics (low aggregation, high photostability, and ...

...is unique compared to the rest of the genome and (2) the sequence is located just upstream of a Protospacer...pyogenes Cas9 variants with varying PAM sequences, 2) the use of Cas9 homologs derived from species other...Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell 163:759–771 . https://doi.... PAMs, respectively (Walton et al., 2020). Characterization of Cas9 from additional bacterial species ...additional Cas9 species PAM sequence may not be characterized Non-SpCas9's bind a variety of PAM sequences.... Furthermore, non-SpCas9's may have other characteristics that make them more useful than SpCas9. For...

...our deposit spreadsheet to complete Step 1. Steps 2 and 3 are usually very easy — our tech transfer team...columns are shown in red in the spreadsheet (Figure 2) and are described in Table 1. Some cells have drop-down...Filling out and sending the deposit spreadsheet Figure 2: Example of the deposit spreadsheet showing the first... Answer “What does this plasmid do?” in <200 characters. A descriptive purpose focuses on the practical...

... 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...marker expression, and viability. Gates are the parameters the machine uses to differentiate between variations...bit of theoretical background on the different parameters that your gating strategy will be based upon.... and Area Within FSC and SSC, there are three parameters that are being measured: height (H), area (A)...events. You are free to choose any size-related parameter, such as FSC-H, FSC-A, SSC-H, SSC-A, etc. You’...

...target sites for editing. PAM sequences can be from 2-6 base pairs in length and follow the DNA sequence... Eric Perkins Mammalian genomic DNA is not just a 2-D linear molecule snaking through our cells’ nuclei...publicly available metagenomic data including the Tara Oceans Project datasets. This study demonstrates...fused dCas9 to a truncated CIB1 protein from Arabidopsis (CIBN), which they call the LADL Anchor. Using... iterations is the addition of CRY2, another Arabidoposis protein that heterodimerizes with CIBN upon ...

...portion of our careers. In that time we may have only 2-3 formal supervisors that will provide guidance and...be challenging – is advice and guidance from only 2 people enough?  For many years I’ve been organizing...lighthearted, the article does highlight key characteristics of a good mentor: Yoda image courtesy...