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We narrowed to 810 results for: MAL

Showing: 321 - 340 of 810 results
  1. Bioinformatics at Addgene

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    Blog Post
    ...on each other for the entire package to run. The small, portable system provided by a container is sequestered... testing and refinements, we specified a select small number of input file names or filtered the input...of the inputs, such as the file name. We started small with a print statement and tested that the desired...working on one process at a time, and testing a small set of data each time a new process was added, we...
  2. QC Sequencing Technologies at Addgene

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    Blog Post
    ... NGS might not be the best choice for small sample sizes, small regions of interest, or complex sequences.... However, Sanger sequencing remains useful for small sample sizes and shorter regions due in part to ...raw sequencing data to FASTQ format, cleans and normalizes reads using the BBTools suite, and assembles ...
  3. CRISPR 101: Any Base Transversion Editors

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    Blog Post
    ... most common type of base excision repair. But mammals also have an enzyme to remove inosine from DNA:...particular species and more, base editors are getting smaller, more accurate, more efficient, and more diverse...precise, efficient A•T-to-C•G base editing in mammalian cells and embryos. Nature Biotechnology, 42(4)...editors for efficient T-to-G and C-to-G editing in mammalian cells. Nature Biotechnology, 42(10), 1538–1547...
  4. Beyond the Blood Brain Barrier: PHP.B Enters the Central Nervous System Via the LY6A Receptor

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    Blog Post
    ...tracks between strains of mice. This suggests that a small genetic difference between these two mice might ...generating a diverse library of AAV9 mutants that had a small randomized peptide loop inserted in the AAV9 capsid... Project. They focused their search on SNPs and small insertions and deletions (indels) most likely to...about using AAV as a tool for gene expression in mammals. Resources on Addgene.org Search for AAV vectors...
  5. Supporting Reproducibility with a Connected ELN

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    Blog Post
    ... The kind of informal conversations about research that can take place in Slack normally never become ...sources The data entered into an ELN is only ever a small fraction of all data directly relevant to a particular...experiment, series of experiments, or project. ‘Small’ data produced by the research group in spreadsheets...
  6. How to Write a Scientific Review Article

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    Blog Post
    ...from genome-wide studies, as well as results from smaller and more specific studies. I also did not limit...of the iceberg. For writers, this is probably “a normal day.” The way I broke this cycle was to just start...You will also begin to hate the article. This is normal and it means you’re on the right track!  Get feedback...deacetylases in B-and T-cell development, physiology and malignancy." Genes & cancer 6.5-6 (2015): 184. PubMed PMID...
  7. CRISPR 101: Validating Your Genome Edit

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    Blog Post
    ...knock-in size ratio below 10:1. Small knock-ins Is your knock-in edit small, with only a few base pairs modified...A method applicable to knock-ins both large and small is a software very similar to TIDE known as TIDER...for specific homology-directed repair events and small edits (Brinkman, et al., NAR). It works just like...
  8. Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus

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    Blog Post
    ...vector like AAV? By breaking the transgene into smaller pieces. Reassembly of fragmented genomes is the...lower than expression of wild-type dystrophin in normal muscles, mice treated with the overlapping split...and acceptor sequences were tested and with the optimal sequences, the group achieved ~80% of muscle cells...application (Trapani et al., 2015).  Expression of smaller fragments of the transgene have been seen with ...
  9. The Advances Behind The World's First Personalized CRISPR Treatment

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    Blog Post
    ...first paper reporting genome editing by CRISPR in mammalian cells appeared in 2013 (Cong et al., 2013; reviewed...to work on DNA instead. However, they retained a small amount of RNA-editing activity, which can cause ...but its effectiveness varied depending on the chromosomal location, sequence context, and cell type. Multiple...Hille, L. T., Ogul, A. N., Robinson-Garvin, N. A., Small, J. C., McCague, S., Burke, S. M., Wright, C. M....
  10. CRISPR 101: Homology Directed Repair

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    Blog Post
    ...oligodeoxynucleotides)) are commonly used for smaller modifications (~1-50 base pairs). Small edits require as little as ...ligation of sticky or blunt ends and often introduces small insertions and deletions along the way. HDR uses...double-strand break repair and their potential to induce chromosomal aberrations. Pfeiffer P, et al. Mutagenesis (...
  11. Using Phosphoserine to Study Protein Phosphorylation

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    Blog Post
    ...phosphorylation events in the proteomes of higher mammals (2). How can we begin to understand the importance...the phosphoryl-modification; they are both much smaller, and carry only a single negative charge as opposed...sequencing of prokaryotic lineages revealed a curious anomaly amongst a subset of methanogenic archea: these ...samples on SDS-PAGE gels containing Phos-Tag, a small molecule that binds specifically to phosphate moieties...
  12. Antibodies 101: The Basics of Western Blotting

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    Blog Post
    ...its primary structure and separated by size with smaller molecules moving more quickly through the matrix...for SDS-PAGE, measure the protein content and normalize to ensure equivalent loading. Denature the samples...at the bottom of the gel and separate by size. Smaller proteins encounter less resistance in the acrylamide...separation, vary the acrylamide content of the gel. For small proteins, use a higher percentage of acrylamide ...
  13. CRISPR 101: Off-Target Effects

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    Blog Post
    ...typically 20 nts and the genomic locus must often be proximal to a PAM site for DNA cutting to occur. The RNA...experiment, at least one will likely be useful. Optimal gRNA selection With some CRISPR experiments, there...can reduce off-targets at a fundamental level. Optimal Cas selection Once you’ve selected a gRNA with ... off-target confounding your results should be minimal. For example, if your goal is to generate a knock-out...
  14. PiggyBac-ing Through the Genome Editing Field

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    Blog Post
    ...successfully used for genetic manipulation in mammalian cells until 2005 [1]. Like other transposons, ...to efficiently perform genetic manipulations in animal models or cells lines, including stem cells, without...specific, and has not been shown to cause genomic abnormalities when expressed. This lack of off-target effects...27075976. 4.  X. Li, E. R. Burnight, A. L. Cooney, N. Malani, T. Brady, J. D. Sander, J. Staber, S. J. Wheelan...
  15. RNA Interference in Plant Biology: New Tools for an Old Favorite

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    Blog Post
    ...trigger in the mammalian research community. Fortunately, many tools for shRNA in mammals are available...enzymes are involved in the biogenesis of different small RNAs (Figure 1). These siRNAs can exert their gene...posts Resources on Addgene.org Find tools for mammalian RNAi Browse all plasmids for RNAi Check out Addgene's...
  16. Mycoplasma Contamination: Where Does It Come From and How to Prevent It

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    Blog Post
    ...Mycoplasmas are considered to be one of the simplest and smallest bacteria. The absence of a rigid cell wall makes...microscope, the mycoplasma DNA appears in the form of small clusters, apart from the cellular DNA. Fluorescent... a very good practice for prevention. Prepare a small quantity of media stock that would be used for the...) Sensitive Assay for Mycoplasma Detection in Mammalian Cell Culture. Anal Chem 84:4227–4232 . https:/...
  17. Technologies Enabled by NanoLuc® Luciferase

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    Blog Post
    ...perform whole-animal imaging, however NLuc’s emission peak is at 460 nm thus making it non-optimal for in vivo...absorbance of heme-containing proteins in whole animals, light at or above 600 nm wavelengths should be...issues as traditional luciferases emit at near optimal wavelengths (Fluc emits at 610 nm) and do not require...
  18. Lambda Red: A Homologous Recombination-based Technique for Genetic Engineering

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    Blog Post
    ...-selectable sequences, point mutations or other small base pair changes, and the addition of protein tags....   This setup can be also be used to modify chromosomal genes. After the initial editing event, the defective... system is best suited for editing bacterial chromosomal DNA because it is easy to remove the recombineering...Knockout/Knockin Plasmids to Disrupt Genes in Mammalian Cells CRISPR Methods for Bacterial Genome Engineering...
  19. Typing CRISPR Systems

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    ... only recently that researchers began building optimal plasmids to harness the power of Class 1 systems...effector and is able to target and edit RNA in mammalian cells. Type IV Type IV is the middle child (like.... Cas9 was the first to be adapted for use in mammalian systems, with the landmark publication in 2013... of DNA. The V-F variants, which are among the smallest Cas enzymes (400 to 700 amino acids), use a Cas14...
  20. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

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    Blog Post
    ...FP-protein fusions (FP-chimeras) is that they undergo normal protein turnover. FP-chimeras are continuously ... Threonine 203 to Histidine (T203H) (1). wtGFP normally contains a mixed population of neutral (protonated...30°C and this is not ideal for experiments in mammalian cells. Dendra2 is another monomeric PA-FP which...microscopy (PALM, also FPALM) (11). For PALM, a small area containing inactive PA-FPs is photoactivated...
Showing: 321 - 340 of 810 results