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Enter a sequence to perform a BLAST-based search of Addgene plasmid sequence databases.

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We narrowed to 1,052 results for: sta

Showing: 781 - 800 of 1052 results

  1. Golden Gate Assembly Upgrades: More Fragments, Faster Assembly, and Higher Fidelity

    Type
    Blog Post
    ...each step. Factors such as enzyme efficiency, stability, and buffer compatibility have placed practical...et al. at NEB (1, 2). While T4 DNA Ligase, the mainstay of most biotechnological cloning efforts for over...for all 3- and 4-base overhang sequences under standard reaction conditions. This data set allows quantitation...

  2. Plasmids 101: Protein Expression

    Type
    Blog Post
    ...glycosylation and acetylation to create functional, stable proteins. Protein expression refers to the second...can express proteins both transiently or through stable cell lines. Both methods produce high protein yields...adapted to fermentation allowing for large-scale, stable production of proteins. In general yeast expression...

  3. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    ...cloning rings to isolate colonies of antibiotic-resistant cells; during the time required to grease, place... below) is simple: After obtaining antibiotic-resistant colonies, use a Sharpie to circle 12-48 individual...cells did not seem to form discrete colonies but constantly migrated away from each other. In our experience...

  4. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    ...locus-specific gRNAs Step 4: Select for puromycin resistant cells Step 5: PCR amplify and sequence the locus...into the FBL locus. Upon sequencing of puromycin resistant clones, they found that 80% and 50% of clones ...using the nuclease Cpf1. Since Cpf1 cuts in a staggered pattern, it is thought to be ideal for HDR-independent...

  5. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...could be combined effectively (3). In Figure 1 for instance, the DBD of the fusion protein shown is still ...to yeast transformation protocols also helped establish the yeast two-hybrid (Y2H) system as a robust ...subset based on a cellular compartment or growth stage). A prey library can either be applied to the bait...

  6. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    ...before investing time and resources into making a stable cell line. When expressing a viral construct transiently... its broad host range, and increased particle stability; however, the protein is cytotoxic precluding ...specific experiments being performed and you should understand the guidelines for using such reagents as outlined...

  7. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...efficiency is at least in part due to La binding and stabilizing the 3’ end of the pegRNA, which is not well protected...pegRNAs (epegRNAs), which add a structural motif to stabilize the 3’ end of the pegRNA. As research has just... research; however, microglia are notoriously resistant to viral transduction. Through in vivo screening...

  8. New Acoustic Reporter Genes: Ultrasound Imaging of Gene Expression

    Type
    Blog Post
    ...protein control. We also added the toxin-antitoxin stability cassette Axe-Txe so that the plasmid could be ...cells gave drastic improvement in expression. Stabilizing the gvpA transcript with WPRE reduced the number...days. GVs should be visible under phase contrast starting day 2 after transfection.     Fig: 2: Expressing...

  9. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    ...drops itself neatly into that gap. Consider, for instance, a disease like cystic fibrosis, which can be ... this new approach proved a challenge. The old standby of CRE/flp didn’t work – it turns out these tyrosine... Christopher A. Vakulskas, John A. Walker II, Anastasia P. Kadina, Adrianna E. Zepeda, Kevin Holden, Jonathan...

  10. CRISPR-based Genome Editing Systems For Editing in E. coli

    Type
    Blog Post
    ...alternative versus plasmid-based expression which can be unstable and lack precise control over copy number. MUCICAT... 1-11 copies of integrated genes. We obtained a stable and high-yielding strain which had 6 times the ...faster construction or reprogramming of cells to understand the genetic and biochemical mechanisms behind...

  11. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    ...NGS-based amplicon sequencing. To use the tool, you start by uploading your Sanger sequencing files, either...useful measure for those who are interested in understanding how many of the contributing indels are likely...CRISPR analysis tool is part of Synthego’s full stack Genome Engineering Solutions, which also includes...

  13. Advanced Uses of Cre-lox and Flp-FRT - A Neuroscientist’s View

    Type
    Blog Post
    ...DNA’, is derived from the yeast S. cerevisiae (Gronostajski and Sadowski 1985). The FLP system works in ...needed for later experiments (cellular tracking for instance) can be controlled with FLP-FRT. For further temporal...19396156. PubMed Central PMCID: PMC3655711. 4. Gronostajski, R M, and P D Sadowski. 1985. “The FLP Recombinase...

  14. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...screening experiments as well). Therefore, if you’ve started down the CRISPR path, you’ve probably got a gene...insert this region into a pCAG-EGXXFP plasmid using standard cloning techniques (The XX indicates where your... them in mouse embryos. When researchers first started using CRISPR/Cas9 for genome editing in mice, they...

  15. Important Considerations in Optogenetics Behavioral Experiments

    Type
    Blog Post
    ...study is an excellent example in which the authors started with a brain region that was known to be generally...steep learning curve and many opportunities for mistakes. The easiest way to learn optogenetics is straight...shared with all. Due to the versatility and standardization of genetic tools in the laboratory mouse, most...

  16. Plasmids 101: The Promoter Region – Let's Go!

    Type
    Blog Post
    ...of its parts. Elements such as the antibiotic resistance gene and the origin of replication are necessary...means exhaustive, but should be a good place to start when trying to pick your perfect promoter! Eukaryotic... gene Although this list is a great place to start, the tables above do not delve into the tissue or...

  17. Chemogenetics Guide

    Type
    Guide
    ...they also retain the ability to be activated by standard optogenetics workflows (such as laser or LED illumination...Planning Your Chemogenetics Experiment If you're starting to plan your chemogenetics experiment, here are...

  19. Evolution of Lab Techniques

    Type
    Blog Post
    ...amplification process, rather than just at the end as in standard PCR. This allows the researcher to compare different... interest to create a physical map of the DNA. Starting in the 1970s, restriction mapping was commonly...techniques we use now may soon become obsolete. For instance, the popular genome-engineering tool, CRISPR, ...

  20. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    ...for at least six months. Future directions and obstacles to clinical CRISPR editing Given the success of...diseases to be treated clinically with CRISPR. Many obstacles remain for CRISPR gene editing, but given the ...Sharp PA, Zhang F. In vivo genome editing using Staphylococcus aureus Cas9. Nature. 2015 Apr 9;520(7546);186...
Showing: 781 - 800 of 1052 results