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We narrowed to 712 results for: BASE

Showing: 281 - 300 of 712 results

  1. Plasmids 101: 5 factors to help you choose the right cloning method

    Type
    Blog Post
    ... you. Traditional restriction enzyme and ligase based cloning is a relatively inexpensive way to get the...a try. This method, developed by Invitrogen and based on lambda phage integration sequences, relies on...inserts a breeze. Among them are some of the homology based methods, like Gibson or SLIC, since one can pool...

  2. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    ...viewed more as a guideline, since they are typically based on the copy number of the empty plasmid backbone...matrix could clog the matrix when performing filter based plasmid prepping. Be sure to stay within the recommended...for each plasmid/strain combination individually, based on determining the cell density or by harvesting...

  3. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    ...two broad categories, prokaryotic or eukaryotic, based on the functional elements they contain. Plasmid...polyadenylation? The role of the terminator, a sequence-based element, is to define the end of a transcriptional...terminators are not usually employed in plasmid-based expression systems, so these will not be detailed...

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

    Type
    Blog Post
    ...know it’s there? For this task, you can use PCR based detection, DNA staining, fluorescent tagging, agar...and cost-effective (Sung, 2006). The sensor cell-based method uses specialized cells that can detect the...from different biotech companies which are enzyme-based mycoplasma detection kits. These kits detect mycoplasma...YJ, Hong JT, Chung YB, Lee CK, Song S (2006) PCR-based detection of mycoplasma species. J Microbiol .44...

  6. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...Target cells Varies based on your experiment Culture media for target cells Varies based on your experiment...the oligos below. Do not change the ends; these bases are important for cloning the oligos into the pLKO...experiment Materials for assay Varies based on your experiment Detailed protocols for preparing polybrene, protamine...your cells. However, you should optimize the time based on your cell line and assay: Assay Days post-infection...of pLKO.1 TRC-cloning vector. The vector is 8901 base pairs total, and the stuffer insert is shown in ...

  7. PCR Applications for Copy Number Variation Assays

    Type
    Blog Post
    ... in this blog post, we're going to focus on PCR-based applications (qPCR and ddPCR.) qPCR Quantitative.... Quantitative analysis of copy number variants based on real-time LightCycler PCR. Curr Protoc Hum Genet...genetic elements can be determined by SYBR-Green-based quantitative real-time PCR. J Microbiol Methods....sample with known copy number set as the expression baseline. Copy number is then determined by comparing results...

  8. Fluorescent Tagging of Endogenous Genes with SapTrap

    Type
    Blog Post
    ...lab developed a polymerase chain reaction (PCR)-based system that allows users to generate a plasmid containing...Masato Kanemaki's lab has developed a CRISPR/Cas-based system for tagging endogenous proteins with an auxin-inducible...Schmid-Burgk, Jonathan L., et al. “CRISPaint allows modular base-specific gene tagging using a ligase-4-dependent...

  9. Google Forums Round Up: First Impressions of NgAgo

    Type
    Blog Post
    ...experiences with the new method and look for advice/tips. Based on the discussions in the forums, it appeared that...majority of the respondents (70%) used a lipofectamine-based method to deliver NgAgo ingredients into the cell.... Overall, the real take-home message thus far, based on survey respondents "Final Verdict", is that more...

  11. The Open Repository of CRISPR Screens: CRISPR Screen Data in One Place

    Type
    Blog Post
    ...in order to increase the accessibility of CRISPR-based phenotypic screen datasets. Original published CRISPR...demonstrated the feasibility and power of CRISPR-based library screens in cell lines from humans and other...? The Open Repository of CRISPR Screens (ORCS) database is a new freely available resource developed by...which can also be used to search the entire CRISPR database for particular screens of interest.   Figure...screen data included in the BioGRID ORCS CRISPR database are encouraged to contact BioGRID curators at ...

  14. New CRISPR Tools: Cas7-11 and PASTE

    Type
    Blog Post
    ...would represent a major step forward for CRISPR-based RNA editing - and further experiments strongly supported...CRISPR could cut DNA, the second generation could do base editing, and with the third generation came Prime...ability to now make any small changes, including base edits, insertions, and deletions. But CRISPR’s uses...deletions and small insertions, leaving a several kilobase-sized gap in the gene editing field. The AbuGoot...now developed PASTE, which can “drag and drop” kilobases’ worth of DNA with CRISPR-driven precision into...

  15. Hot Plasmids: Summer 2025

    Type
    Blog Post
    ...label based on antibody-like sensors) changes that. Developed by the Arnold Lab, ATLAS is based on an ...synapses, and cytoskeleton components) and in a FRET-based biosensor for cAMP dynamics. The mGold2 variants...

  16. What’s in Your Tube?: A Quick Guide to Measuring DNA by Spectrophotometry

    Type
    Blog Post
    ...the most popular options, so most of this post is based on reading results from a NanoDrop. The principles...calculate a sample’s nucleic acid concentration based on the sample’s absorption at 260 nm (A260). ...use a complementary assay, such as a fluorescence based assay or running an agarose gel. In the case of ...

  17. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    ...number of ways, from small edits that change single base pairs, to chromosomal inversions and large deletions...1 CRISPR systems make up the majority of CRISPR-based immune systems in bacteria and archaea. Class 1 ...deletions of up to hundreds or even thousands of base pairs, stopping either at boundaries defined by ...originally suspected that Cas3 moved a couple of kilobases down the strand of DNA, Cascade caught up to it...

  18. Career Insights: Technical Support Specialist

    Type
    Blog Post
    ..., technical support is often covered by the lab based team as part of their general job duties. Bigger...etc. Technical support roles are primarily office based, and email/phone are the most popular ways to communicate... pressure which are often found in sales or lab-based roles in industry. What to look forward to in a ...answers for customers, internal resources and databases are scrutinized, and this information is enriched...

  19. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    ... the very beginning of the third chromosome, ten base pairs in length. An emerging family of sequencing...increase the expression of a gene that is millions of base pairs away. (For details on DNA compaction and the...parsed for spCas9 PAM sites (NGG) and then filtered based on two main criteria: no TTTTT allowed (this is ...here with permission. Quilt is a collaborative database for genomics. In this article, Quilt CTO Aneesh...is CTO at Quilt Data. Quilt is a collaborative database. Aneesh's research interests span machine learning...

  20. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...common to use many preys with a single bait in Y2H-based screens. In this way, proteins that interact with...proteome of a particular cell type, or a subset based on a cellular compartment or growth stage). A prey...protein-protein interactions in mammalian cells based on protein splicing. Anal Chem. 2001; 73:2516-2521... Including a prey-only control also provides a baseline level of reporter activation. Vary the expression...
Showing: 281 - 300 of 712 results