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We narrowed to 736 results for: RAN-1

Showing: 681 - 700 of 736 results

  1. SciComm: Taking Science to Elementary and Middle Schools

    Type
    Blog Post
    ...can make this fun activity relate to STEM.” Figure 1 shows the activities we had for my school’s Math &...olds in a single evening. Grades K-5 cover a broad range of attention spans, skill levels, and STEM exposure...aspects of talking to 8th graders was the broad range of questions I received. I gave my presentation ...

  2. Mesothelioma - Causes, Symptoms, and Treatment

    Type
    Blog Post
    ...blood test for HMGB1, the high mobility group box 1 protein. Normally localized to the nucleus, if the...of diagnosis) is about 35 years, but it usually ranges from 20-50 years and sometimes longer. There is...be Used to Study Cancer Drug Resistance Plasmid Grant to Aid Cervical Cancer Screening in Ghana Additional...

  3. The Challenges of Cell Culture

    Type
    Blog Post
    ...testing. You can find Nick on LinkedIn. References 1. Skloot, Rebecca, and Bahni Turpin. The immortal life...cell line contamination: Epithelial Mesenchymal Transition Is Required for Acquisition of Anoikis Resistance... to carry our independent quality control for a range of testing, from animal doping to forensics, DNA...

  4. Four Base Editing Reporters to Monitor and Enrich Editing in Real-time

    Type
    Blog Post
    ...complex. So how exactly does this work? Figure 1: APOBEC- and Cas9-mediated editing (ACE) reporter ... affecting protein translation of different reporter proteins. Protein translation requires the start ...compared to CRISPR/Cas9 edits that rely on double strand DNA breaks. Due to this inefficiency it is crucial...constitutively expressed to identify successfully transfected cells. C-to-T editing by the APOBEC-Cas9 editosome...that results in the restoration of fluorescence. Transient reporter for editing enrichment (TREE) The Wang...Wang and Brafman lab developed a transient reporter for editing enrichment (TREE) to purify single base... to cells segregated solely by a reporter of transfection, which only report the efficiency of plasmid...

  5. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    ...therapies to clinical trials in 2021. Figure 1: Schematic of the human MECP2 locus and MECP2 protein...References: Amir RE, Van den Veyver IB, Wan M, Tran CQ, Francke U, Zoghbi HY (1999) Rett syndrome is caused...collection of molecular biology tools so that both veteran and new Rett researchers can find the reagents ...maturation (Tillotson & Bird, 2019) and is a global transcriptional regulator of thousands of genes. Studies have...have suggested roles in transcriptional repression, activity dependent de-repression, chromatin remodeling...groove of AT-rich DNA sequences to regulate transcription (Baker et al., 2013). Mutations that affect ...gene editing, MECP2 reactivation, RNA editing, RNA trans-splicing, and protein replacement to reverse the...

  6. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    ...levels of DnaA to get the job done.   Figure 1: DNA melting by DnaA and IHF is the first step of ...the proteins cooperatively can separate the DNA strands. Created with BioRender.com. Once DnaA breaks...it recruits helicases to continue unwinding the strands. This creates two replication forks that move in...elongation phase, a DNA polymerase synthesizes a new DNA strand at each replication fork. Termination sequences...act as a feedback mechanism to reduce further transcription. Replication is also regulated by the balance... concentration is controlled at the level of translation. RepZ mRNA must fold into a “pseudoknot” held...intramolecular base pairs for the protein to be translated. An antisense RNA called Inc, also encoded by...

  7. Filming Science Videos in the Age of Social Distancing

    Type
    Blog Post
    ... over again. Here’s that process!   Figure 1: The Design-Build-Test-Learn cycle adapted for video...looking, I found an inexpensive HDMI transmitter that could transmit a signal between a wall. There was ... things first was getting that wireless HDMI transmitter/receiver, to beam the camera’s image to my remote...then set up a monitor connected to the wireless transmitter and an iPad in the office next door (connected...

  8. 5 Reasons to Use Reddit for Science Communication

    Type
    Blog Post
    ...first foray into Reddit. Here are some reasons why: 1. Reddit has a large and active community of people... general advice on how to meet people, how to transition out of academia, and other science career topics.... Although I didn’t start answering until 5pm, frankly, I was typing far before five. Fortunately I have...

  9. General Transfection

    Type
    Protocol
    ...cells were transfected using 1:1, 1:2, 1:3 and 1:6 µg of pRosetta :µg of PEI. The 1:2 and 1:3 ratios provided...18.9 18.9 1:2 18.9 37.8 1:3 18.9 56.7 1:4 18.9 75.6 1:5 18.9 94.5 1:6 18.9 113.4 Gently add the diluted PEI...possible range of ratios to test: Ratio of DNA:PEI Amount of DNA (μg) Volume of 1 mg/mL PEI (μL) 1:1 18.9 .... Thawed aliquots should be discarded after 1–2 months. 1 mg/mL polyethylenimine, linear MW 25,000 Da ...stable. Dilute 1:3 (µg DNA:µg PEI) in 500 µL total of OptiPro SFM (per 10 cm plate). 56.7 µL of 1 mg/mL PEI...for viral production) Day 1: Transfect Cells Day 2 (am): 18 h post transfection - Remove media, replace ...prepared, transfect cells with a fluorescent plasmid using a variety of ratios. Check the cells 1–2 days ...

  10. Antibodies 101: Beyond Surface Labeling

    Type
    Blog Post
    ...and biotin plasmids at Addgene!     Figure 1: Indirect detection of target markers can be achieved...can be motifs within transmembrane proteins, such as receptors, or post-translational modifications on those...diffuse through the cell membrane freely, while others require a permeabilized membrane to enter cells. Depending...label intranuclear proteins of interest, you can label cytosolic proteins alongside the intranuclear ones...Depending on the application, they can be membrane-permeant or membrane-impermeant, and the cells of interest...viability dyes are membrane-impermeant and can only enter cells with compromised plasma membranes — i.e., dead...conjugates are commercially available for a wide range of fluorophores, allowing for great flexibility ...

  11. Transfection for Recombinant Antibodies

    Type
    Protocol
    ...determined for your sample. Typical ratios may range from 1:1 to 1:6. Add the diluted PEI-MAX to the diluted...solution at -20 °C. 1 mg/mL PEI-MAX Add 1 g of PEI-MAX powder to 900 mL deionized water in a 1 L bottle and ...to mix. Add 450 µL of 1 mg/mL PEI-MAX to the second tube of 6 mL BCD TFX (for a 1 mg/mL stock solution ...antibody At 168 hours (1 week) post-transfection, harvest the antibody. Transfer the HEK293 cells and media...February 18, 2022 Workflow Timeline Day 1: Seed cells Day 2: Transfect cells Day 3-6: Feed cells Day 7: Harvest...at -20 °C. Procedure Section 1: Seeding cells The day prior to transfecting, seed a 108 mL culture of HEK293.... Cap the tubes and incubate for 1 h in the 37 °C bead bath. Transfer the PEI-MAX and recombinant antibody...

  12. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

    Type
    Blog Post
    ...mycoplasma can even escape 0.2-micron filters. Figure 1: Contamination! Tissue culture media often have indicators..., and effective for wide range of mycoplasma. These methods cannot guarantee complete removal of mycoplasma...the media after autoclaving hence, there is no assurance of the serum being contamination free. The aerosols...commonly infecting mycoplasmas are used. A broad range of primers encompasses almost all the infecting ...filters, for media filtration. There is no 100% assurance that this process can avoid contamination but ...

  13. Antibody Guide

    Type
    Guide
    ...and placed in a buffer. Antibody Structure Figure 1: Structure of an antibody A standard antibody is made... together to form a “Y” shape, as shown in Figure 1. The two arms of the Y structure are responsible for...proteins by size. Transfer proteins from the gel to a membrane. Incubate the membrane with an unconjugated...block the membrane to remove excess antibody and prevent unwanted binding. Incubate the membrane with a ...Wash and block the membrane. Activate the HRP with a substrate kit. Image the membrane using either X-ray...hybridoma cells into plasmids. Plasmids are then transfected into cells and induced to express antibodies....variable regions, shown in light blue and light orange, that bind to a specific antigen, triggering an...

  14. Protocol - Bacterial Transformation

    Type
    Protocol
    ...get more colonies if you use 1 μl of a 1:5 or 1:10 dilution rather than 1 μl directly.... 20-30 mins. Heat shock each transformation tube by placing the bottom 1/2 to 2/3 of the tube into a 42...and then (optional) incubate in 37°C incubator. Mix 1 - 5 μl of DNA (usually 10 pg - 100 ng) into 20-50 ...Introduction Transformation is the process by which foreign DNA is introduced into a cell. Transformation of bacteria... more easily transformed and that will help to maintain the plasmid without rearrangement of the plasmid...prepared for optimal transformation efficiencies upon thawing. For the highest transformation efficiency, we...Pro-Tips Commercial competent cells range significantly in their transformation efficiency. The lowest efficiency...

  15. FLEx Technology and Optogenetics: Flipping the switch on gene expression with high spatial and temporal resolution

    Type
    Blog Post
    ...halorhodopsins to silence neuronal activity (Figure 1) (Wiegert et al., 2017). How to get an opsin into...flow of electric charges across cell membranes and maintain membrane potential in response to visible light...across the axon's membrane: once a certain number of positive ions crosses the cell membrane, a threshold ...rhodopsins enable ions to flow across the axon’s membrane, thereby controlling neural activity (Bernstein...the opsin into the correct orientation to be transcribed (Figure 2) (Sharma and Zhu, 2014). How does FLEx... optogenetic FLEx vectors in combination with transgenic animals or rAAVs expressing Cre under a specific...the mechanisms of sleep and anesthesia. In Cre transgenic mice, they targeted the dopamine (DA) neurons...

  16. A Guide to Getting Started in Undergrad Research

    Type
    Blog Post
    ...a short-term position (1-2 years) Often a gap-year position taken to transition to the next career stage...investigator. In basic science (as opposed to translational, biomedical, or applied sciences), people conduct...don’t tell us everything there is to know is a strange and exciting revelation. Science in any field can...investigator (PI) Runs the lab Writes a lot of grants Varying degrees of engagement with what happens...

  17. Molecular Cloning Techniques

    Type
    Guide
    ...more in our Restriction Cloning blog post . Figure 1: Restriction enzyme cloning of your gene of interest...by certain bacteria and archaea, cleave double-stranded DNA (dsDNA) at specific sequence sites in the ...into fragments containing precise 5' or 3' single-strand overhangs (sticky ends), or no overhang (blunt ... They also cut specific target sequences, which range from four to 13 base pairs, and produce predictable... with four nicks that bacteria repair after transformation. LIC does not require site-specific recombination...exonuclease until a base is exposed on the single-strand overhang that is complementary to the free nucleotide... is rendered unnecessary. The product may be transformed directly into E. coli , where the normal replication...

  18. Finding Your Perfect Job After University

    Type
    Blog Post
    ...experience in cancer research After graduating with a 2:1 BSc in Molecular Biology (roughly a B average in the...project to validate some genomic changes, so this arrangement was ideal for everyone.  The cutting edge science...young scientists and PhD students to also explore a range of alternative careers outside of acidemia to have...

  19. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...Consortium A.2 Map of pLKO.1 A.3 Related plasmids B. Designing shRNA Oligos for pLKO.1 B.1 Determine the optimal...Order oligos compatible with pLKO.1 C. Cloning shRNA oligos into pLKO.1 C.1 Recommended materials C.2 Annealing... References H.1 Published articles H.2 Web resources I. Appendix I.1 Sequence of pLKO.1 TRC-Cloning Vector...marker encoded in pLKO.1 allows for convenient stable selection. Figure 1 : Map of pLKO.1 containing an shRNA...puromycin should be from 1-10 μg/mL in 1 μg/mL increments. d. Label plates from 1-10 and add appropriate...Appendix I.1. Sequence of pLKO.1 TRC-Cloning Vector Click here to see the sequence of pLKO.1 TRC-cloning...convenience. See “warranty information” in appendix. Table of Contents A. pLKO.1-TRC Cloning Vector A.1 The RNAi...

  20. Lentivirus Production

    Type
    Protocol
    ...ratios of 1:1, 1:2, 1:3 and 1:6. The 1:2 and 1:3 total DNA:PEI μg ratios provided high transfection efficiencies...possible range of ratios to test: Ratio of DNA:PEI μg of DNA μL of 1 mg/mL PEI 1:1 18.9 18.9 1:2 18.9 37.8... 37.8 1:3 18.9 56.7 1:4 18.9 75.6 1:5 18.9 94.5 1:6 18.9 113.4 Gently add the diluted PEI mixture to the...: Monday: Plate 1×10 6 cells in a T75 flask in 15 mL DMEM Complete. Wednesday: Plate 1×10 6 cells in a...packaging cells Day 1 (pm): Transfect packaging cells Day 2 (am): 18 h post-transfection. Remove media, replace...ratio of μg DNA:μg PEI is 1:3 (1000 μL total per 10 cm dish). Using transfer plasmid pHAGE TRE dCas9-KRAB...prepared, transfect cells with a fluorescent plasmid using a variety of ratios. Check the cells 1-2 days ...
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