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We narrowed to 592 results for: cel.2;

Showing: 541 - 560 of 592 results

  3. Save Time with Transient Plant Leaf Transformations

    Type
    Blog Post
    ...GUS-staining is a very useful visual reporter (Figure 2). The GUS enzyme cleaves a substrate, which will then...roots, as functional chloroplasts and leaf specific cell types are missing in roots. The development of transgenic...transformation” usual means that neither a stable cell line or plant is regenerated from the transformed...with the gene of interest is transferred into the cells. But for C. roseus, Agrobacterium infiltration into...fused to a protein of interest to elucidate the subcellular localization of your protein of interest. Find...Intrinsic GUS-like activities in seed plants." Plant cell reports 9.1 (1990): 1-5. PubMed PMID: 24226366. ...

  4. To Codon Optimize or Not: That is the Question

    Type
    Blog Post
    ...Hoogsten base pairs and wobble pairs.  Figure 2: (A) The codon wheel depicts the 64 codons that encodes...mammalian cells. This was the first codon optimized membrane protein expressed in mammalian cells. In this...sequence. That means there are 61 possible tRNAs. Cells however, may not express all 61 of these tRNAs and... are also optimized for expression in mammalian cells. For example, Henry Lester’s lab optimized the C...Caenorhabditis elegans GluCl ion channel genes for mammalian cells dramatically improves expression levels. Journal...

  5. Plasmids 101: Codon usage bias

    Type
    Blog Post
    ...PMID: 18478103. PubMed Central PMCID: PMC2364656. 2. Dittmar, Kimberly A., et al. "Selective charging ...reasons include: Metabolic pressures - it takes cellular resources to produce tRNAs that recognize different...mismatched translation rates and potential effects on cell growth, even hosts producing alternative tRNAs may...inclusion bodies that will generally segregate with the cell pellet during purification procedures. Even if you...a means to fine-tune gene expression." Molecular cell 59.2 (2015): 149-161. PubMed PMID: 26186290. PubMed...

  6. Protocols for Molecular Biology, Plasmid Cloning, and Viral Preps

    Type
    Protocol
    ...BSL-1 and BSL-2 Labs Learn how to best protect yourself when working in BSL-1 and BSL-2 labs Watch the...Two (BSL-1 and BSL-2) Safety measures for laboratories operating at BSL-1 and BSL-2 Watch the Video! Water...Generating Stable Cell Lines with Lentivirus Genomically integrate your DNA sequence into a cell line Isolating...Isolating a Monoclonal Cell Population by Limiting Dilution Generate monoclonal cell lines from a polyclonal...polyclonal pool of stable cells AAV Production in HEK293 Cells Produce adeno-associated virus to deliver your...Extraction Without a Kit Extract RNA from tissues or cells without a commercial kit Plasmid Cloning Protocols...Transfection Introduce plasmid DNA to mammalian cells Lentivirus Production Produce lentivirus with a ...

  7. Course-Based Undergraduate Research Experiences (CUREs)

    Type
    Blog Post
    ...that view (Callahan et al., 2022).     Fig. 2: Students in an MDH CURE replaced select residues ...identifying new genes required for stem and progenitor cell maintenance in planarians in Developmental Biology... his teaching style, with the former offering excellent training in improvisation and the latter making...junction complex is required for stem and progenitor cell maintenance in planarians. Developmental Biology...

  8. Transferable Skills Guide: Career Planning Resources

    Type
    Blog Post
    ...American Society for Cell Biology (ASCB) or the American Chemical Society (ACS) are just 2 examples of organizations...for the position. This post on Business Insider nicely summarizes good job fair practice. Career pages..., there is an etiquette that you should follow, nicely summarized in this post on Forbes.com, and here...

  9. Protocol - Bacterial Transformation

    Type
    Protocol
    ...each transformation tube by placing the bottom 1/2 to 2/3 of the tube into a 42°C water bath for 30-60 ...depending on the competent cells you are using). Put the tubes back on ice for 2 min. Add 250-1,000 μl LB...the competent cells in your hand instead of on ice Reduce step 4 from 20 - 30 mins to 2 mins on ice before...commonly referred to as 'competent cells.' Many companies sell competent cells, which come frozen and are prepared...that came with your competent cells. Pro-Tips Commercial competent cells range significantly in their ...SOC media Competent cells DNA you'd like to transform Procedure Take competent cells out of -80°C and thaw...use electro-competent cells. Instead of relying on the heat-shock to cause the cells to take up the DNA,...

  10. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...vapors. Workflow Timeline Day 1: Antigen Coating Day 2: Blocking Day 3: Primary antibody incubation Day 4...with 96-well plates 1–10 µL single channel pipette 2–20 µL single channel pipette 20–200 µL single channel...serial dilutions of the purified antigen as follows: 2 ng/µL : Add 100 µL of 20 ng/µL stock into 900 µL PBS...microfuge tube and vortex. 1 ng/µL : Add 450 µL of 2 ng/µL stock into 450 uL PBS in a microfuge tube and...37 °C for 30 min , or overnight at 4 °C . Section 2: Block the plate Prepare the wash buffer (0.05% Tween...incubate on a microplate shaker set to 400 rpm for 2 h at room temperature or overnight at 4 °C . Section...

  11. Immunocytochemistry

    Type
    Protocol
    ...HeLa cells per well. Allow the HeLa cells to grow to the desired density before labeling. Section 2: Fixing...coated coverslips HeLa cells 24-well plate 4% Paraformaldehyde 5 mg/mL 4′,6-diamidino-2-phenylindole (DAPI...Fluorescent microscope 0.5–10 µL single channel pipette 2–20 µL single channel pipette 20–200 µL single channel...of the diluted antibody to the wells and incubate 2 h at RT . Remove the primary antibody and dispose ...detect antigens in cells. Here we describe the basic steps for fixing and labeling cells in culture with ...2022 Workflow Timeline Day 1: Seed cells Day 3-4: Fix and label cells Equipment Pipette controller Pipette... Seeding cells Place a sterile poly-D-lysine coated coverslip in each well of a 24-well cell culture treated...

  12. Bioinformatics at Addgene

    Type
    Blog Post
    ...the necessary steps to perform:      Fig. 2: Quick-stepping to a FASTA file   With our outline...mission and our own, as both address the need to accelerate science using different approaches. We were also...evolved into a single, organized pipeline and accelerated the processing of our sequencing data. However...

  13. Protocol - How to Inoculate a Bacterial Culture

    Type
    Protocol
    ...mL glass bottle: 4 g NaCl 4 g Tryptone 2 g Yeast Extract and dH 2 O to 400 mL Note: If your lab has pre-mixed... to do a mini-prep you will usually want to start 2 mL in a falcon tube, but for larger preps you might...might want to use as much as a liter of LB in a 2 L Erlenmeyer flask. Using a sterile pipette tip or toothpick...bacterial cell. Large plasmids usually have a low copy number (approximately one or two copies per cell) and... can be present in large numbers, 50 or more per cell and have a high copy number. High copy number plasmids...increase this to 350 - 400 rpm to obtain a higher cell density. Reference Page Top Index...

  16. CRISPR Library Amplification

    Type
    Protocol
    ...times are limited. Day 2: Harvest cells and purify DNA (Estimated time 3-4 hours) Cells should be harvested... recover, set up overnight growth (Estimated time 2-3 hours) Transformation should be performed at the... Tubes should contain a total of 5 mL (3 mL SOC + 2 mL transformed Endura from two separate transformations...has been absorbed by the agar. This usually takes 1-2 minutes. Critical Be careful not to rip or shred the...incubation if needed. Place 100 mL sterile LB at 4 ℃. Day 2 (morning) Before beginning, prechill at least four...pellet. The total weight of each pellet should be ~1-2 g. Pro-Tip Make sure to weigh the empty tube beforehand...electrocompetent cells (Default: 4 tubes of Endura Duos, Lucigen, 60242-1) Alternatives include Stbl4 cells or other...

  17. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    ...infection of cultured cells.” Nature 339 (1989). 487–488. PubMed PMID: 2725683. 2. Miller, C.J., Kassem...Love your cells and they'll love you back A successful lentiviral infection begins with your cells, both ...both your packaging cells and your target cells. Poor tissue culture practices can lead to low titers and... promote cell growth. Overgrown cultures stimulate quiescence, the resting phase of the cell cycle; quiescent...quiescent cells do not take up nucleic acid or express transgenes as well as actively dividing cells resulting...reaching 108 cells per ml before the media becomes cloudy. Mycoplasma compete with host cells for nutrients... your producer cells with fresh medium. Daily medium exchanges help alleviate cellular stresses associated...

  18. Proximity Labeling: A Powerful Tool for Protein Complex Purification and Proteomic Mapping

    Type
    Blog Post
    ...approximately 2-4 million proteins per cubic micron in bacteria, yeast, and mammalian cells (Milo, 2013)...about RapID! EXCELL: Proximity labeling for the detection of cell-cell interactions EXCELL is an enzyme-mediated...enzyme-mediated proximity cell labeling strategy to detect cell-cell interactions in vivo (Ge et al., 2019...et al. "Enzyme-Mediated Intercellular Proximity Labeling for Detecting Cell–Cell Interactions." Journal...a bait protein fused to BirA* in cells and then incubate those cells with exogenous biotin. After a few...peptide (LPETG). In EXCELL, the mgSrtA variant can be displayed on the surface of a cell of interest, where...-protein interactions, and even more recently cell-cell interactions. If you’re looking to identify interactions...

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

    Type
    Blog Post
    ...positive cells by the number of only mCherry positive cells (Martin et al., 2019). Figure 2: Editing...restoring mCherry expression. Cells can then be sorted using Fluorescence-activated cell sorting (FACS) and the...easily determined by comparing cells expressing only eGFP expression to cells expressing eGFP and mCherry...expressed in cells. mCherry is constitutively expressed to identify successfully transfected cells. C-to-T ...TREE) to purify single base edited cells without the need for single cell isolation and downstream sequencing...fluorescence and cell sorting using FACS which could limit their use across different cell types and systems...sgRNA targeting the GFPACG to the cells. As expected, only the cells that contained the cytidine base ...

  20. The Fluorescent Vegetables in Aptamer Soup

    Type
    Blog Post
    ...of 3,5-difluoro-4-hydroxybenzylidene-imidazolinone-2-oxime (DFHO), a fluorophore modified from DsRed. DFHO...challenging to track intracellularly, but aptamers permit the study of intracellular RNA dynamics. Aptamers...RNA aptamers are used to track cellular processes by transforming cells with plasmids encoding aptamer-tagged... been used to investigate intracellular viral DNA dynamics in neuronal cells (Nilaratanakul et al., 2017...be used to simultaneously target cellular RNA to specific intracellular locations using Cas9 and track ...and later introducing the small molecule into the cell. The fluorescence that is generated allows for qualitative...). DFHBI is membrane permeable and non toxic to cells. Because it rapidly photobleaches, DFHBI can only...
Showing: 541 - 560 of 592 results