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We narrowed to 492 results for: ara-2

Showing: 201 - 220 of 492 results

  1. Deep Dive: qPCR

    Type
    Blog Post
    ...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...

  2. Selecting Your Plasmid Purification Kit

    Type
    Blog Post
    ... 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...

  3. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

    Type
    Blog Post
    ...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...

  4. Viral Vectors 101: Inverted Terminal Repeats

    Type
    Blog Post
    ...ITR, which can either be “flip” or “flop” (Figure 2). Additionally, the ITR contains a 4-nucleotide Rep...site for Rep proteins (Savvy et al. 2013). Figure 2: AAV2 wild type ITR in flip and flop orientation. ...Packaging, and Potency of AAV Vectors. Microorganisms, 12(2), 310. https://doi.org/10.3390/microorganisms1202031038399714..., B-B’, and C-C’) that give the feature its characteristic T-shape. The arrangement of the B-B’ and C-...

  5. Transferable Skills Guide: Cross-team Communication

    Type
    Blog Post
    ...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...

  6. Plasmids 101: CcdB - The Toxic Key to Efficient Cloning

    Type
    Blog Post
    ...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...

  7. What's Your Organism? Expanding Genomic Tools via the NSF EDGE Program

    Type
    Blog Post
    ...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, ...

  8. Plasmids 101: Plasmid Incompatibility

    Type
    Blog Post
    ...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...

  9. Antibodies 101: Reading a Flow Plot

    Type
    Blog Post
    ...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...

  10. Retrograde AAV viral preps

    Type
    Collection
    ...physically separate nuclear dTomato, Cre-dependent Calcium sensor Ting 83899 pAAV-mDlx-GCaMP6f-Fishell-2 mDlx...final viral vector preparation. In some cases, in vivo testing of these viral preparations was performed by...Addgene is producing ready-to-use viral vector preparations in a retrograde serotype from the Alla Karpova...David Schaffer laboratories. These viral vector preparations were produced with the rAAV2-retro helper plasmid...-Ef1a-mCherry-IRES-Cre EF1a Cre and physically-separate mCherry expression Recombinases Deisseroth 105540...pAAV-EF1a-mCherry-IRES-Flpo EF1a Flpo and physically-separate mCherry expression Recombinases Deisseroth 55636...GCaMP6f-P2A-nls-dTomato Syn GCaMP6f and physically separate nuclear dTomato Calcium sensor Ting 51083 AAV-...

  11. Scientific Peer-review: Providing Critical and Kind Feedback and Advocating for Open Science

    Type
    Blog Post
    ...-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...

  12. Hot Plasmids: Winter 2025

    Type
    Blog Post
    ...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...

  13. Hot Plasmids: Fall 2024

    Type
    Blog Post
    ...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...

  14. Plasmids 101: Broad Host Range Plasmids

    Type
    Blog Post
    ...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...

  15. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    ...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 ...

  16. The PAM Requirement and Expanding CRISPR Beyond SpCas9

    Type
    Blog Post
    ...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...

  17. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

    Type
    Blog Post
    ...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...

  18. Antibodies 101: Introduction to Gating in Flow Cytometry

    Type
    Blog Post
    ... 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’...

  19. Hot Plasmids - September 2019 - CRISPR, Auxin Inducible Degradation, MerMAID Channelrhodopsins, and Genome Looping

    Type
    Blog Post
    ...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 ...

  20. "What Makes a Good Mentor?" and 6 More FAQs About Science Mentoring

    Type
    Blog Post
    ...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...
Showing: 201 - 220 of 492 results