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We narrowed to 604 results for: abo.2

Showing: 541 - 560 of 604 results

  1. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    ...al (Nature. 1988 Jul 28;334(6180):320-5.) PubMed. 2. A segment of the 5' nontranslated region of encephalomyocarditis...genes above with your gene or genes of interest. Some of the plasmids listed in the table above (and their... an internal region of the mRNA. In the figure above, you can see that the IRES element acts as another... first discovered in picornaviruses, are short (about 20 amino acids) and produce equimolar levels of ...

  2. New Optimized Genome-wide CRISPRko, CRISPRi, and CRISPRa Libraries

    Type
    Blog Post
    ... PMID: 25184501. PubMed Central PMCID: MC4262738. 2. Doench, John G., et al. "Optimized sgRNA design to...Read this blog post about CRISPR Kinome Libraries available at Addgene Learn about the advantages of pooled...large-scale functional screens Read this blog post about genome-wide screening using CRISPR Resources on...libraries for the mouse genome! Other Resources Read about CRISPRi and CRISPRa from the NIH E-newsletter ...

  3. Negotiating Work and Life: How to Find the Joy

    Type
    Blog Post
    ...work. Nobody wins, and you need to make a change. #2 Thou shalt sometimes put work ahead of life Sometimes...on the Society for Laboratory Automation and Screening (SLAS) Electronic Laboratory Neighborhood.   Work...worry less about how they will handle an emergency and this will make you much more confident about their ...

  4. Plasmids 101: Codon usage bias

    Type
    Blog Post
    ...PMID: 18478103. PubMed Central PMCID: PMC2364656. 2. Dittmar, Kimberly A., et al. "Selective charging ...that lead to these preferences and what can we do about them? Read on to find out!   Why do organisms have...understood, but some possible reasons include: Metabolic pressures - it takes cellular resources to produce...amino acid I’d like to encode,” but, as described above, not every region of a protein should necessarily...Effects on transcription (remember, it’s not all about translation - e.g. codon choice may interrupt transcription...

  5. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...agarose concentrations (e.g., 2 g of agarose in 100 mL of TAE will make a 2% gel). Mix agarose powder with...concentration of approximately 0.2-0.5 μg/mL (usually about 2-3 μl of lab stock solution per 100 mL gel). EtBr...gels are commonly used in concentrations of 0.7% to 2% depending on the size of bands needed to be separated...allows you to gauge how far the DNA has migrated; 2) it contains a high percentage of glycerol that increases...cool down to about 50 °C (about when you can comfortably keep your hand on the flask), about 5 mins. Optional... of the tip of the pipette into the buffer just above the well. Very slowly and steadily, push the sample... of the way down the gel. A typical run time is about 1-1.5 hours, depending on the gel concentration ...

  6. Design Tips for Prime Editing

    Type
    Blog Post
    ...inhibit editing (Anzalone et al., 2019). Figure 2: Starting lengths for optimizing your pegRNA primer... primer binding site, starting with a length of about 13 nt (Anzalone et al., 2019). Primer binding sites...of reverse transcriptase template, starting with about 10–16 nucleotides. For longer templates, it is even.... Test multiple nick sites, starting with sites about 50 bp upstream and downstream from the prime editing...

  7. 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...Generating a transgenic plant takes even longer, is more labor intensive, inefficient, and often not reproducible... region is transferred into the plant. In the laboratory we can utilize this natural process by replacing...system. The vectors mentioned in this blog post above are ready to use vectors, but with the MoClo system...

  8. 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...usage bias Read more about plasmids in our Plasmids 101 blog series Learn more about trends in synthetic... out Addgene's Molecular Biology Reference Read about genetic code expansion Browse our plasmid collection...

  9. Hot Plasmids - August 2020

    Type
    Blog Post
    ...based antiviral method to degrade RNA from SARS-CoV-2 sequences and live influenza A virus. It was demonstrated... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here.  Here's... a cell. The Goedhart lab, Gadella lab, and collaborators have an ongoing effort to improve genetically..., that are optimized for excitation wavelengths above 1,000 nm and enable two-color imaging with red fluorescence...

  10. Bioinformatics at Addgene

    Type
    Blog Post
    ...the necessary steps to perform:      Fig. 2: Quick-stepping to a FASTA file   With our outline...much information as possible from depositing laboratories and associated publications, but our quality...the process. Another design choice we appreciated about Nextflow was how effortless it was for us to switch...interface. In our next post, we’ll share our story about how we used Nextflow Tower to augment our pipeline...

  11. Transferable Skills Guide: Career Planning Resources

    Type
    Blog Post
    ...) or the American Chemical Society (ACS) are just 2 examples of organizations that hold large, annual ...or postdoc forever, and you need to be thinking about what comes next. To that end, one of the most important...personal connection; the importance of which will be elaborated on later. As for what job fairs to consider, ...the specific position he or she originally called about, you can keep in touch and reach back out when you...

  12. Bacterial Toxin-antitoxin Systems as Molecular Biology Tools

    Type
    Blog Post
    ...mazF gene and a gene for Zeocin resistance (Figure 2). This MazF-ZeoR cassette is flanked by two direct... more about CcdB, the toxic key to efficient cloning Read all Plasmids 101 blog posts Read about knockout...with the other toxin-antitoxin systems described above, one part of the pair is more stable than the other...

  13. High Complexity Golden Gate Assembly with NEB’s DAD

    Type
    Blog Post
    ...rules are: (1) don’t use the same overhang twice; (2) avoid palindromes; (3) no overhangs with the same...cycling. The Pros and Cons Researchers used the tools above to complete a 52-piece assembly of a 40 kb T7 phage... help from DAD. The 35-piece assembly mentioned above had a remarkably high 71% fidelity, while the 52...

  14. Antibody Validation Using the Indirect ELISA Method

    Type
    Protocol
    ...section 1, step 2 is a suggested starting point. If your unknown sample’s absorbance falls above the range ...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...

  15. Kit Free RNA Extraction

    Type
    Protocol
    ...RNAzol®, QIAzol® (for Protocol Option #2) Water-saturated Phenol 2 M Sodium Acetate pH 4 Chloroform/Isoamyl... reagent section above for the recipe). If using TRIzol®, jump down to the Option #2 - TRIzol® Protocol....5% (wt/vol) N-laurosylsarcosine (Sarkosyl) 0.1 M 2-mercaptoethanol TRIzol® or similar product such as...following sequentially to 1 mL of lysate: Add 0.1 mL of 2 M sodium acetate (pH 4.0), mix thoroughly by inversion...aliquots of it and storing those in -80°C. Option #2 - TRIzol® Protocol Homogenize or lyse tissues or cells...by hand for 10 seconds. Incubate the sample(s) for 2-3 minutes on ice and centrifuge for 15 minutes at ... working with the volatile reagents in the list above. Procedure Option #1 - Solution D Protocol Before...

  16. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    ...rate of 10-55% - the PASTE system.     Fig. 2: Schematic of programmable gene insertion with PASTE.... Eugene Koonin and Kira Makarova, longtime collaborators of Omar and Jonathan and experts in CRISPR evolution...

  17. Hot Plasmids - November 2023

    Type
    Blog Post
    ... vivo and in vitro applications.   Figure 2: Modular features of DIRECTED particles where the ...section of the Addgene antibody page, and learn more about the whole-brain reference atlas efforts, including...

  18. Viral Vectors 101: Preparing Pooled Libraries

    Type
    Blog Post
    ...replicate, then purify the DNA.     Figure 2: A pooled library page with protocols boxed in red...ordering a pre-made library, you don’t need to worry about amplifying and producing lentivirus. But pre-made...

  19. Immunocytochemistry

    Type
    Protocol
    ...Fluorescent microscope 0.5–10 µL single channel pipette 2–20 µL single channel pipette 20–200 µL single channel... plate 4% Paraformaldehyde 5 mg/mL 4′,6-diamidino-2-phenylindole (DAPI) Bovine serum albumin (BSA) Triton...grow to the desired density before labeling. Section 2: Fixing and permeabilizing cells Gently aspirate the...of the diluted antibody to the wells and incubate 2 h at RT . Remove the primary antibody and dispose ...water Microscope slide Anti-fade mounting medium Laboratory wipes 15 mL conical tubes 50 mL conical tubes...paraformaldehyde and follow your institution's laboratory safety guidelines for disposing of waste in the...remove the coverslip. Blot the coverslip with a laboratory wipe to remove excess liquid. Add 1 drop of anti-fade...

  20. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ... glacial acetic acid 57 mL of dH 2 O Store Solution III at 4°C Grow 2 mL overnight cultures from single...Resuspension buffer Denaturing solution Renaturing solution 2 mg/mL RNase A TE or water-saturated phenol-chloroform...100 μL of cold Solution I. Vortex the solution for 2 min or until all bacteria are fully resuspended. Add...pipetting or carefully pouring. Optional: Add 5 μL of 2 mg/mL RNase A to the supernatant in the new tube and... to the recovered aqueous DNA layer. Repeat steps 2-4. Note: Phenol-chloroform is a hazardous waste - ...: Ethanol Precipitation To your DNA solution, add 2-2.5 volumes 95% or 100% ethanol and 1/10 volume of... supernatant out of the tube if you are worried about losing the pellet. Dry with vacuum or by inverting...
Showing: 541 - 560 of 604 results