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  1. Single-cell tracking of lineage and identity with CellTag

    Type
    Blog Post
    ...with reprogramming outcomes (Biddy et al., 2018). Tracking lineage and cell identity at the single-cell level...intestine, demonstrating its utility in long-term tracking in vivo (Guo et al., 2019). Key takeaways of CellTag...cell-indexing tool that combines scRNA-seq and lineage tracking to track the dynamics of gene expression and clonal...

  2. Bricking Science: Portraying Scientific Reality Through LEGO

    Type
    Blog Post
    ...contributed by guest blogger, Dalila Cunha de Oliveira. Bricking Science is an idea built, literally, 'brick-by-brick...Running Smoothly and "Gnome-Proof" This is why the “Bricking Science” YouTube channel was born. The name came...feet), to connect people with science. And so, “Bricking Science” was born. We chose the stop motion format...to follow protocols, guidelines, and standards. Bricking science reminded us of our inner children who ...in LEGO form, feel free to share with us at BrickingScience@gmail.com. Who knows, your story might just...student in Hematology from Brazil and co-founder of Bricking Science.      Additional Resources on the Addgene... bricking science...

  4. CRISPR Plasmids - Prime Edit

    Type
    Collection
    ...pegRNA + nicking sgRNA BbsI No Vermilion Norbert Perrimon 164423 pHSG1C3 Mammalian U6 pegRNA + nicking sgRNA...promoter, the type of gRNA (e.g., pegRNA, epegRNA, nicking sgRNA), cloning enzyme, selectable marker, and ...mRFP1 David Liu 140448 QPM-sgR (pTaU3) Plant TaU3 nicking sgRNA Eps3I + NcoI No Caixia Gao 141081 pYPQ141D-peg...Weber 172717 pnsgRNA Bacteria J23119 (BBa_J23119) nicking sgRNA No Tilmann Weber 173220 pPBT-peRNA_GG-Puro...sgRNA-mCherry Mammalian, AAV hU6 + H1 pegRNA + nicking sgRNA No mCherry Hyongbum Kim 177180 pDAS12069_...

  5. Zhang Lab CRISPR Page

    Type
    Collection
    ...Cre-dependent and constitutively expressing Cas9 knockin mice (Platt et al ., Cell 2014). In these mice ...applications. Full references are below. The Cas9 knockin mouse can be purchased from Jackson Labs: Cre-dependent... for the mouse Rosa26 locus; Used to make Cas9 knockin mouse Vectors for cancer modeling 60224 : sgRNAs...10.1038/nbt.2647. Epub 2013 Jul 21. PubMed . Double nicking by RNA-guided CRISPR Cas9 for enhanced genome editing...plasmids AAV Plasmids with Cas9 Mouse CRISPR-Cas9 knockin mice for genome editing and cancer modeling. Platt...

  6. Bacterial Expression Systems

    Type
    Collection
    ... Genome Engineering Although overexpressing or knocking down a gene can tell you much about its function...crystallography, you’ll likely need to purify it. Luckily, many proteins can be functionally expressed in...Growth of strain USO hisB- pyrF- rpoZ- on plates lacking histidine and containing 3-AT Scot Wolfe Bacterial...- pyrF- rpoZ- , this promotes growth on plates lacking His and containing 3-AT. FLIParaF.Ec-250n 18855...

  7. CRISPR Guide

    Type
    Collection
    ...dynamic tracking of repetitive and non-repetitive genomic loci, chromosome painting, and RNA tracking in living...inhibit CRISPR by a variety of mechanisms, including blocking crRNA loading or PAM recognition; inhibiting Cas9...sites is sufficient to repress transcription by blocking initiation. More robust transcriptional repression...proteins that target RNA instead of DNA enables tracking of RNA in live cells, including orthogonal RNA...Smith, S. B., Meadows, S. K., Roberts, B. S., Mackiewicz, M., Mendenhall, E. M., & Myers, R. M. (2015)...

  8. CRISPR Plasmids - Cascade-Cas3

    Type
    Collection
    ...is a unique member of the CRISPR family. After nicking the target DNA, Cas3 initiates single-strand DNA...target DNA using a gRNA and recruits Cas3. After nicking the non-targeted strand, Cas3 starts unraveling...

  9. Immunocytochemistry

    Type
    Protocol
    ...PBS on a rocking platform. Permeabilize cells for 10 min at room temperature ( RT ) on a rocking platform...PBS on a rocking platform. Section 3: Labeling with antibody Block for 20 min at RT on a rocking platform...platform in 500 µL blocking buffer. Remove the blocking buffer and dispose of it in an appropriate waste container...Equipment Pipette controller Pipette tips and pipettes Rocking platform Tweezers Fluorescent microscope 0.5–10...buffer: Dilute 20 µL of Triton X-100 in 10 mL PBS. Blocking buffer: Dilute 0.5 g BSA and 30 µL Triton X-100...container. Wash 3x for 5 min in 500 µL PBS on a rocking platform. Dilute the fluorescently-labeled secondary...container. Wash 3x for 5 min in 500 µL PBS on a rocking platform. (Optional) Counterstain nuclei with 500...

  10. Western Blot

    Type
    Protocol
    ...information specific to your antibody, such as ideal blocking buffer and optimal antibody concentrations. Consider...When the run is complete, select Done. Section 5: Blocking Prepare 1X TBST as follows: 25 mL of 20X TBS 2.5...20 472.5 mL of deionized water Mix well Prepare blocking buffer as follows: Dilute 5% w/v non-fat milk ...milk into 100 mL of 1X TBST. Pro-Tip The ideal blocking buffer will vary between antibodies. Refer to ...1X TBST, protein side up. Block the membrane in blocking buffer for 1 h at RT on a shaking platform. Wash...primary antibody to the desired concentration in blocking buffer. Pro-Tip The optimal concentration will...membrane overnight in primary antibody at 4 °C on a rocking platform. Pro-Tip Primary antibody incubation can...

  11. Lentivirus Plasmids

    Type
    Collection
    ... ORF expression, multicistronic design enables tracking of viral transduction. Also see plasmid 46948 ...inducible knockdown, multicistronic design enables tracking of viral transduction. See article for more similar...

  12. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ...Microcentrifuge Electrophoresis chamber Power supply Rocking platform Fume hood Metal spatula Razor blade Plastic...deionized water for 5 min with gentle agitation on a rocking platform. Pour off the water in the sink. Add 20...and incubate for 1 h with gentle agitation on a rocking platform. Pour off the SimplyBlue SafeStain in ...and incubate for 1 h with gentle agitation on a rocking platform. Pour off the water in the sink. Add 100...and incubate for 1 h with gentle agitation on a rocking platform. Pour off the water in the sink. Take ...

  13. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...Workflow Timeline Day 1: Antigen Coating Day 2: Blocking Day 3: Primary antibody incubation Day 4: Secondary...aspirate the wash buffer from the wells. Prepare the blocking buffer (1% BSA in PBS) as follows: Add 250 mg ...mix. Using a multichannel pipette, add 200 µL of blocking buffer to each well. Cover the plate with a plate...

  14. Fluorescent Protein Guide: Biosensors

    Type
    Collection
    ... novel analytical method for in vivo phosphate tracking. FEBS Lett. 2006 Oct 30. 580(25):5885-93. Wolf...Glutamate iGluSnFR glutamate sensor with improved trafficking to the plasma membrane Acute disruption of the...

  17. Institute for Protein Innovation

    Type
    Collection
    ...ligand-binding pocket Bind specific subunit pairs, blocking ligand binding and receptor function Figure courtesy...

  18. FlyCRISPR

    Type
    Collection
    ...replace genes or other genomic sequence with an attP docking site. Contains the visible marker 3xP3-DsRed. (...

  19. General Transfection

    Type
    Protocol
    ...DNA. Add the diluted PEI dropwise while gently flicking the diluted DNA tube. Incubate the mixture 15–...DMEM complete. Incubate the cells 24–48 h before checking for protein expression. Sample Data Legend: Lenti-X...

  20. Neurodegeneration Plasmid Collection

    Type
    Collection
    ...CMV ALS Richard Kriwacki 205856 CL20_mEGFP_MFF_ERBB4 ERBB4 GFP CMV ALS Richard Kriwacki 205928 CL20_mEGFP_SSBP2...Richard Kriwacki 205953 CL20_mEGFP_WASL_VPS13D VPS13D GFP CMV Spinocerebellar Ataxia Richard Kriwacki 206069...
Showing: 1 - 20 of 330 results