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  2. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    Blog Post
    ...non-functional 𝛽-galactosidase. Complementing this mutation by expressing this deleted portion, for example...custom primers are especially helpful in verifying mutations that wouldn't be reached by sequencing from either...

  3. Hot Plasmids - May 2022

    Type
    Blog Post
    ...long sequences with reduced risks of unwanted mutations in mammalian cells. Long sequences traditionally...traditionally have higher risks of off-target mutations. These plasmids contain catalytically inactive dCas9 and...

  4. Plasmids 101: CcdB - The Toxic Key to Efficient Cloning

    Type
    Blog Post
    ...gene. One such strain is DB3.1, which contains a mutant version of DNA gyrase (gyrA462) that is resistant... al. “F plasmid CcdB killer protein: ccdB gene mutants coding for non-cytotoxic proteins which retain ...

  5. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    ...vector with half the size of the ssAAV genome has a mutation in the terminal resolution site (TRS) to form ...genome with wild-type ITRs at the both ends and mutated ITR at the center of symmetry. After uncoating ...

  8. Fluorescent Proteins 101: Introduction to FRET

    Type
    Blog Post
    ...orientation limiting FRET efficiency, circular permutation (Baird et al., 1999) of the fluorescent protein...References 1. Baird, Geoffrey S., et al. "Circular permutation and receptor insertion within green fluorescent...

  9. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

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    Blog Post
    ...indicated in the “Species Other” column. Relevant Mutations Indicate the amino acid change as well as any...functional consequences of that change. For example, “Mutated aspartate 123 to Lysine (D123K), lowers biosensor...

  10. Rabies and Neuronal Tracing

    Type
    Blog Post
    ...with a deletion-mutant rabies virus. Retrograde neuronal tracing with a deletion-mutant rabies virus. Pubmed...

  12. Progress Towards a PAM-Free CRISPR

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    Blog Post
    ... if you want to knock out a gene, a frameshift mutation introduced anywhere within the early coding region... hand, if you are trying to knock-in a patient mutation or need to make a precise amino acid change, there...

  14. Design Tips for Prime Editing

    Type
    Blog Post
    ...with BioRender.com. Add silent mutations near point mutations to create 3-base (or longer) tracts...

  15. CRISPR in the Clinic

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    Blog Post
    ...co-injection of Cas9 and a single-guide RNA targeting the mutant allele. When all was said and done, the researchers...targeted to precisely edit with the goal of ‘fixing’ mutations or introducing a desired edit, but this particular...

  16. Open Resources and Plasmid Tools For Studying C. elegans

    Type
    Blog Post
    ...additional SapTrap plasmids used for introducing point mutations that was published in a microPublication (Schwartz... E. M. "SapTrap vectors for introducing point mutations with unc- 119 + selection." MicroPublication 1...

  19. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    ...due to its endA1 recA1 relA1 genotype, as these mutations improve plasmid stability and yield. If a plasmid...using older plates could increase plasmid loss or mutation. Let it incubate Increasing the incubation time...

  21. Tips for Using BLAST to Verify Plasmids

    Type
    Blog Post
    ...BLAST (blastx) search to check for expected ORFs, mutations or truncations. A primary advantage of blastx ...database. Another benefit is that a frameshift mutation present in the ORF is readily apparent when viewing...

  22. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    ...outlined below (or emission wavelenght range).  By mutating GFP, the variants blue FP (BFP), cyan FP (CFP)...breakdown of GFP, it's variants, and their relevant mutations, check out Marcy's post from last week on GFP....

  25. CRISPR 101: Ribonucleoprotein (RNP) Delivery

    Type
    Blog Post
    ...et al. 2018). This was accomplished using a Cas9 mutant (Cas9M1C/C80S) that harbors two surface exposed...disulfide-activated ligands. Due to the concerns of off-target mutations and transgene integration in crops, delivery of...

  26. Plasmids 101: Colony PCR

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    Blog Post
    ... but won’t tell you if your products have any mutations.  There are lots of different cloning strategies...expected sized PCR product doesn’t mean there aren’t mutations in your insert. Make sure to submit multiple positive...

  27. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    ...the deletion reduces the risk of insertional mutagenesis by limiting the interaction between viral LTR...potential for oncogenesis through insertional mutagenesis. These risks are largely based upon the vector...

  28. Mesothelioma - Causes, Symptoms, and Treatment

    Type
    Blog Post
    ...inflammation, which can cause mesothelial cells to mutate and ultimately form cancerous tumors.  This process...For example, multiple studies show that certain mutations of the BAP1 gene play a role in susceptibility...

  29. Hot Plasmids Spring 2024

    Type
    Blog Post
    ...HaloTag, Figure 4), they determined that an A122D mutation in the rhodopsin domain conferred several new ...potentials of fast-spiking neurons. The same A-to-D mutation in the related voltage indicator Ace2N-mNeon also...

  31. Finding nucleic acids with SHERLOCK and DETECTR

    Type
    Blog Post
    ...determine the presence and quantity of DNA, RNA or a mutation of interest. Together SHERLOCK and DETECTR demonstrate...SHERLOCK could also detect low-frequency cancer mutations from cell-free DNA fragments as well as health-related...

  32. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    ... HEPN domains in other proteins. As with Cas9, mutating key residues in the Cas13a molecule results in...Zika virus, genotype human DNA, identify tumor mutations within cell-free genomic DNA, and detect COVID...

  33. CRISPR 101: Engineering the Plant Genome Using CRISPR/Cas9

    Type
    Blog Post
    ...2013) Plant genome editing made easy: targeted mutagenesis in model and crop plants using the CRISPR/Cas...Rice Blast Resistance by CRISPR/Cas9-Targeted Mutagenesis of the ERF Transcription Factor Gene OsERF922...

  34. Hot Biosensors 2022: Year-End Roundup

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    Blog Post
    ...biosensors are generated by inserting a circularly-permuted GFP into an intracellular loop of various G-protein-coupled...around 70 mM and 100 mM, respectively. Introducing mutations and using potassium-binding protein homologues...

  35. Fluorescent Proteins 101: When GFP lets you down

    Type
    Blog Post
    ...mEGFP. This variant was made by introducing two mutations (F64L, S65T) in the original jellyfish GFP (AvGFP...with improved brightness (Tsien, 1998). Another mutation (A206K) is necessary to generate the strictly ...

  37. CRISPR 101: Epigenetics and Editing the Epigenome

    Type
    Blog Post
    ... spiroplasma (M. Sss1) (termed MQ1.) The Q147L mutation improves methylation kinetics such that cytosine...fused LSD1 to x-Cas9(3.7). x-Cas9(3.7) is a SpCas9 mutant that exhibits increased PAM flexibility, and this...

  38. Important Considerations When Using AAVs

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    Blog Post
    ...them. There are suspected cases of insertional mutagenesis in humans (1) and AAVs are non-enveloped viruses... Russell, David W. "AAV vectors, insertional mutagenesis, and cancer." Molecular Therapy 15.10 (2007):...

  39. Quick Guide to All Things Lentivirus

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    Blog Post
    ...active area of research due to the possibility that mutation or recombination with human retroviruses could...Efficient gene transfer Potential for insertional mutagenesis: Replication-incompetent lentiviruses with human...

  40. Live and Let Dye: Self-Labeling Protein Tags

    Type
    Blog Post
    ... most often consist of an enzyme that has been mutated to block its catalytic activity and instead become... later work showed that introducing a Leu28Cys mutation in eDHFR does lead to covalent binding of the ...

  41. 9 tips for a successful postdoctoral experience

    Type
    Blog Post
    ...with senior investigators. Discussions can reveal mutual scientific interests, complementary perspectives...research to search committees, and can lead to mutually beneficial collaborations. 7. You cannot get too...

  47. CRISPR Cheat Sheet

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    Blog Post
    ...multiple genes in a cell population. The resulting mutant cells are screened for phenotypes of interest. ...

  48. New Viral Vectors - Summer 2024

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    ...viral service with multiple serotypes pAAV-hSyn-GRAB_OTmut AAV9 Biosensors (Control) Li New viral service...

  49. What's New in CRISPR - May 2019

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    Blog Post
    .... Thus, they engineered two CBE variants with mutations in the rat APOBEC1 that decreases the number of...
Showing: 151 - 200 of 398 results