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We narrowed to 801 results for: Ran

Showing: 481 - 500 of 801 results

  1. Screening for Successful Genome Editing with Digital PCR

    Type
    Blog Post
    Published
    March 30, 2017, 2:30 p.m.
    ...edit. Since exploitation of NHEJ to generate translational reading frame-altering indels for “functional... an unedited site distal tothe predicted double strand break, and a NHEJ/ “drop-off” probe designed to...to bind directly at the predicted double strand break site (Figure 2A). The single molecule resolution...PubMed PMID: 26914317. 4. Mock, U. et al. mRNA transfection of a novel TAL effector nuclease (TALEN) facilitates...

  2. Plasmids 101: Methylation and Restriction Enzymes

    Type
    Blog Post
    Published
    June 30, 2016, 2:30 p.m.
    ...DNA. The other component of these systems, methyltransferases, methylate DNA at particular sequences to...restriction modification systems containing methyltransferases that add methyl groups to specific DNA sequences...overlaps with the XbaI cut site (depicted as an orange line) because the restriction site is followed ...plasmids with no methylation (there are no methyltransferases in the PCR reaction). The template plasmid...

  3. REPLACR Mutagenesis: Replacing In Vitro Recombination Methods

    Type
    Blog Post
    Published
    Feb. 10, 2016, 3:30 p.m.
    ... and the nonmethylated PCR products are then transformed into E. coli to undergo nick repair. This method...containing the desired mutations, to amplify both strands of a plasmid template. In addition to containing...template plasmid, these PCR products are directly transformed into bacteria expressing recombineering proteins...fine, especially using plasmids in the 10-12 kb range. REPLACR-mutagenesis also requires making your own...

  4. 3 Challenges in Plant Synthetic Biology

    Type
    Blog Post
    Published
    July 22, 2014, 5:46 p.m.
    ...how they started their company here. The first transgenic plant was engineered over 30 years ago, but plant... still in its infancy. A long timeline from transformation to testing and a lack of well-characterized...s made even more difficult by the fact that transformation methods have also been patented. In order to.... We will also be working with open source, transformation technologies like those pioneered by Ewen Mullins...

  5. Troubleshooting Your Plasmid Cloning Experiment

    Type
    Blog Post
    Published
    Sept. 24, 2019, 12:55 p.m.
    ...the specific reaction you are running. Transformation Transform a few microliters of your ligation reaction...The PCR could fail to produce a product, the transformation may not result in any cells, or all the colonies...fragments If you don’t get any positive clone, try transforming the ligation mix in different bacterial strains...

  6. Rosella: A Fluorescent pH-Biosensor for Studying Autophagy

    Type
    Blog Post
    Published
    April 13, 2017, 2:30 p.m.
    ...Autophagosomes are relatively easy to identify: double-membraned structures containing undigested cytoplasmic contents...stands for protein) associates with autophagosomal membranes, so tagging it with GFP allows for tracking the...emission spectra. Component Excitation Emission pH Range DsRed.T3 488, 543*, 568 nm 587 nm ~4.9 - ...mechanisms behind autophagy by tracking what’s being transported to the yeast vacuole (i.e. cytosol, mitochondria...

  7. RNA Extraction Without A Kit

    Type
    Blog Post
    Published
    March 24, 2020, 1:15 p.m.
    ...expense, and less of being left with a bunch of random solutions when all the spin columns run out. In... inherently not as stable as DNA- it is single stranded and its ribose groups are susceptible to hydrolysis...effectively isolate intact RNA, a fast, strong protein denaturant would be required- something that broke down ...Chirgwin and colleagues showed that a strong protein denaturant, guanidinium thiocyanate, did just this (Chirgwin...

  8. Celebrating 15 Years of Scientific Sharing

    Type
    Blog Post
    Published
    Jan. 14, 2019, 2:08 p.m.
    ...technical support Our customer and technical service brands continue to be impeccable. We score ~99.9% helpful...lauded by another set of stakeholders - the tech transfer community. In a recent law review article we received... have a UI/UX team. Last time I checked we were ranked as the 21,806th most popular website in the world...pages and excellent work on our external website appearance and functionality are happening and this strengthens...

  9. Summer SciComm: Preprints

    Type
    Blog Post
    Published
    Aug. 16, 2022, 1:15 p.m.
    ...conference poster or talk, or submitting patent or grant applications; to informal settings like lab meetings..., and increase their impact. Open access and transparency Another major benefit of preprints is their ... of listing “Submitted” articles in their CV or grant applications. A number of organizations (such as...be prone to biases or errors, with a lack of transparency in most review processes. Preprints and their...

  10. To Each HIS Own

    Type
    Blog Post
    Published
    March 2, 2023, 2:15 p.m.
    ...and several contaminating proteins (blue, green, orange) flows through the Ni2+-containing resin. The protein...polyhistidine containing contaminating protein (orange). A low imidazole wash buffer disrupts the weak...polyhistidine-metal ion interaction is not disrupted by denaturants (Hochuli, 1988). In some cases, proteins can ...tag and then refolded by gradual removal of the denaturant. Not all proteins will refold once denatured,...

  11. Plasmid-based Recombinant Monoclonal Antibodies: What They Are and Why You Should Be Excited About Them

    Type
    Blog Post
    Published
    May 4, 2021, 1:15 p.m.
    ...chain and light chain for the antibody and when translated into protein will assemble into a fully functional...synthesize the genes that encode those amino acids (Tran et al., 2016). Each antibody would then be tested...P, Frenzel A, Wong J, Koh XY, Eng H-Y, Lane D, Lefranc M-P, Clark M, Dübel S (2018) When monoclonal antibodies...:633–644 . https://doi.org/10.1083/jcb.201409074 Tran NH, Rahman MZ, He L, Xin L, Shan B, Li M (2016) ...

  12. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

    Type
    Blog Post
    Published
    April 6, 2021, 12:30 p.m.
    ...fall within a 10 fold range. Plasmids that don't meet the lower bound of this range are “failed” and removed...mutations, we check to see if they affect the translated amino acid. We also confirm that the species ...2017) Genome-scale CRISPR-Cas9 knockout and transcriptional activation screening. Nat Protoc 12:828–863...

  13. Split Fluorescent Proteins for Studying Protein-Protein Interactions

    Type
    Blog Post
    Published
    May 3, 2022, 1:15 p.m.
    ...fragments are named according to the number of strands in the canonical beta-barrel structure. Most frequently...frequently, the proteins are split between strand 10 and 11 to yield large (1-10) and small (11) fragments...oligomerisation. Eastwood T, Baker K, Brooker H, Frank S, Mulvihill DP. FEBS Lett. 2017 Dan Mulvihill ...shifting tag (FAST) for use with green-yellow or orange-red fluorogen A split fluorescent reporter with...

  14. Making CRISPR Plasmids Using Fragmid

    Type
    Blog Post
    Published
    Aug. 29, 2024, 1:15 p.m.
    ...Gate cloning, perform an exonuclease cleanup, transform, plate, and pick. In short, the Fragmid system...AAV pRDA_889 (ITRs from AAV2) Mammalian Transposon pRDA_757 (Piggybac; constitutive expression...   Drosophilia Plasmid pRDB_053 (for transgenic flies; gypsy insulator sequences; white + selectable... drosophila) Delivery type (lentivirus, AAV, transposon, Plasmid) Destination vector (dropdown options...

  15. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

    Type
    Blog Post
    Published
    May 15, 2018, 12:43 p.m.
    ...expression in E. coli. This operon is under the transcriptional control of a strong constitutive promoter (... required enzymes, including a glutathione S-transferase homolog from a Janthinobacterium sp. Moreover... protein kinase A (PKA) activity at the plasma membrane in living cells using a construct in which TagRFP-T...used in quenching, FRET (Förster resonance energy transfer), and FLIM (fluorescence lifetime imaging microscopy...

  16. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    Published
    July 2, 2020, 12:40 p.m.
    ...sequences that, when bound by transcription factors, can increase a gene’s transcription. Sometimes enhancers ...and proteins that forms chromosomes. These long range DNA interactions are often detected with chromosome...control gRNA or predicted gRNA off-target sites. Long-range chromatin interactions identified with the multiplexed...

  17. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    Published
    May 4, 2017, 2:30 p.m.
    ...production of dsRed E5. This timer predictably transitions from green to red fluorescence (500 to 580 nm... of vesicle fusion and release from the plasma membrane by fusing various vesicle cargoes to FP timer ...fusing Medium-FT to LAMP-2A, a lysosome-associated membrane protein, and following its subcellular localization...whether or not newly synthesized or long-lived transcription factors control gross changes in gene expression...

  18. Hot Plasmids: Winter 2025

    Type
    Blog Post
    Published
    March 11, 2025, 1:15 p.m.
    ...NC: nucleocapsid RNA chaperone, RT: reverse transcriptase. Right: T1 tomato plants that inherited an edited...Cultured rat hippocampal neuron expressing pan-membrane ASAP5. Solid blue circle marks the initiation ...via the Kv2.1 targeting motif as well as a pan-membrane construct without the Kv motif. Altogether, if...kinetics and high responsivity near the resting membrane potential, ASAP5 is a great new option. Find ASAP5...

  19. Starter guide to induced pluripotent stem cells (iPSCs) part 1:  A renaissance in regenerative medicine

    Type
    Blog Post
    Published
    Oct. 30, 2018, 1:04 p.m.
    ...during ESC isolation. Low Chances of post-transplantation rejection when used therapeutically High Low...similar functional utility, iPSCs bypass the transplantation compatibility issues and ethical concerns of... used therapeutically, the chances of post-transplantation rejection are low. Further, as iPSC generation...somatic cell type to another, also known as transdifferentiation.   Many thanks to our guest blogger, Kusumika...

  20. Plasmids 101: 5 factors to help you choose the right cloning method

    Type
    Blog Post
    Published
    Aug. 21, 2018, 12:31 p.m.
    ...would at least be guaranteed a successful result? While there are never any guarantees, some cloning techniques...Resources” section at the end of the post! Speed: Your grant deadline is approaching or your advisor is demanding...your insert, DpnI digest away the templates, and transform the PCR products into E. coli. Ligases within ...
Showing: 481 - 500 of 801 results