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Showing: 321 - 330 of 595 results
  1. Plasmids 101: Terminators and PolyA signals

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    ...due to the presence of additional helper sequences (2-3). As alluded to above, termination and polyadenylation...and translation processes as well.   Figure 2: The conserved eukaryotic polyadenylation signal directs...Plasmids designed to express genes in a given host cell type are generally broken down into two broad categories...purpose and mechanism of polyadenylation vary across cell types, but polyadenylation generally serves to promote... thought that poly(A)s are used to control the cellular concentration of regulatory RNAs and may additionally...additionally act as a quality control mechanism to rid the cell of mis-folded RNAs. Eukaryotic termination and polyadenylation...signal by using in vitro analyses. Molecular and Cellular Biology. 1992. PMID: 1333042. PubMed Central PMCID...
  2. 28 Hot Plasmid Technologies from 2015

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    ... Zetsche et al., Nat Biotechnol. 2015 Feb 2;33(2):139-42. doi: 10.1038/nbt.3149. Light-inducible ...and on your favourite signalling pathways (Figure 2). They are all available now at Addgene. 1 Lungu...20;19(4):507-17. 10.1016/j.chembiol.2012.02.006. 2 Guntas et al., Proc Natl Acad Sci USA 2015 Jan 6; ...Nihongaki et al., Chemistry and Biology. 2015 Feb 19; 22(2):169-74. High efficiency cloning for enzyme screening...inducible, gene knockdown with shRNA or shRNA-miR30, (2) tamoxifen-inducible CreERT2 gene deletion, (3) simultaneous...different fluorescent proteins, 8 constitutive promoters, 2 includible promoters, 3 polyA terminators and various...of plasmids required for each input/output system; 2) removing all chemically inducible promoters; 3) improving...
  3. 7 Tips to Secure a STEAM Internship This Summer

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    Blog Post
    ...and scholarships in different subject areas!    2. Resume and cover letter know-how  Tailor your resume...4. Letters of recommendation wisdom  Reach out to 2-3 individuals in your professional circle to vouch...college. Prior to March 2020, internships were a fiercely competitive endeavor. Due to the COVID-19 pandemic...
  4. Form Your Own Peer Mentoring Group: A How-To Guide for Scientists

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    Blog Post
    ...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...do once you are in a mentoring relationship.  Celebrate your successes and support each other as you transition...
  5. 5 Tips for Troubleshooting Viral Transductions

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    ...Central PMCID: PMC3760253. 2. Rohwer, Forest. "Global phage diversity." Cell 113.2 (2003): 141. PubMed ... digest and compare it to some reliable controls. 2. Viral stocks may be sensitive to freeze thaw cycles...contaminating your transduction with the packaging cells. Packaging cell debris can be removed either by filtration... you can reduce the volume of cell culture medium on the packaging cells immediately following transfection...efficiency in cells that are sensitive to the toxicity of polybrene (e.g., hematopoietic or primary cells), and... the packaging cells after 1–3 days of selection. A lower percentage of surviving cells may be the result...adeno-associated virus transduction of cells." American Journal of Physiology-Cell Physiology 291.2 (2006): C386...
  6. Viral Vectors 101: Voltage Indicators

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    ...Indicators GEVIs can be divided up structurally into 2 main groups: voltage sensitive domains and Rhodopsin...depending on your experimental system.   Figure 2: General overview of chemigenetic tools. A genetically...Cellular communication and function depend on potential energy. Cellular activities like action potentials...Indicators As previously mentioned, cellular voltage is measured at the cell membrane, and this voltage is ...endocrine cells. It follows that the specific targeting of these voltage indicators needs to be cell type ...electrical events! Voltage indicators are voltmeters for cells! In this blog we will review how these indicators...related to the electrical impulse of the cardiac cells. To study this phenomenon you will likely want to...
  7. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

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    Blog Post
    ...from GFP and the mutation definitions found in Table 2 may not apply. Table 1: The specific mutations comprising..., V163A EBFP F64L, S65T, Y66H, Y145F   Table 2: Functional role of specific mutations in GFP derivatives...measure, and control molecular signals, cells, and groups of cells in order to understand their activity...Fluorescence-activated cell sorting (FACS): This is a type of flow cytometry that separates mixtures of cells into distinct...Thus, FACS can be used to separate cells expressing GFP from cells that are not. Developmental/transgenic... localization profiles, acting as a biosensor or cell marker, studying protein-protein interactions, visualizing... was non-toxic and could be expressed in living cells, enabling the study of dynamic, physiological processes...
  8. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

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    Blog Post
    ...membrane integrity (Fig. 2). For example, saponin interacts with cholesterol in cell membranes and leaves ... at too high a concentration.      Figure 2: Examples of different permeabilization strategies...holes in the cell membranes of your sample to allow antibodies better access to intracellular targets. If...allowing antibodies into the cell where they could bind to intracellular epitopes. C) Permeabilizing with...method for visualizing molecules of interest within a cell or tissue. In that introduction, we broke down the...structure of your samples, the relationships between cellular components, and the accessibility of your target...cross-linkers work by (as the name suggests) cross-linking cellular proteins. Cross-linking is a chemical reaction...
  9. Google Forums Round Up: First Impressions of NgAgo

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    ... PMID: 27136078. 2. Burgess, Shawn., et al. "Questions about NgAgo." Proteins & Cell (2016). 3. "NgAgo...engineering methods have their own pros and cons (2). The NgAgo DNA guide is inexpensive and can be 5’...degraded by the cellular machinery, leaving a small amount of active NgAgo/ssDNA complex in the cell. As proposed...results of Gao et al 2016 in a letter to Protein & Cell. Update (August 3rd, 2017) THE ORIGINAL NgAgo ARTICLE... guide, it cannot be produced from a plasmid in cells. Additionally, in vitro assembly of NgAgo/ssDNA ...dangerous, non-physiological temperature for mammalian cells and one which diminishes the endonuclease activity...guide. Therefore, researchers must co-transfect cells with 5’-P-ssDNA guides and an NgAgo expression plasmid...
  10. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

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    ...amilCP, tsPurple, and gfasPurple) (Figure 2). Figure 2: Chromoprotein expression from bacteria on... them into your cells, you can simply look for blue colonies to determine which cells have taken up the... that the cellular fitness cost varied greatly between chromoproteins. In some cases, cells containing... mScarlet as the donor in HeLa cells.   Find ShadowR for mammalian cell expression here   Now that you...cloning experiment worked just by looking at the cells on the plate. For example, if your plasmid contains...Here, the chromoprotein is expressed and gives the cell a magenta (amilCP) or purple (tsPurple) color. Though...added which induces expression of the toxin. Any cells that grow after this point will be those that have...
Showing: 321 - 330 of 595 results