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We narrowed to 881 results for: NSI

Showing: 601 - 650 of 881 results

  2. Mouse Modeling, Part 2: Breeding and Crossing Mice

    Type
    Blog Post
    ...Special breeding cases Whether your mouse colony consists of genetically engineered mice with a single mutation...strains that have been crossed, there are many considerations for effectively breeding your mice. Because...

  3. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    ...lack all viral genes, including those that are responsible for integration into host chromosomes, further...to other gene delivery tools that are under consideration, recent advancements in various AAV technologies...

  4. Progress Towards a PAM-Free CRISPR

    Type
    Blog Post
    ...the ‘quality’ of the gRNA sequence, including propensity for off-targets, secondary structures, etc., ...more pan-genome PAM flexibility, you’ll want to consider one of several Cas9s that recognize NG PAMs. The...

  5. 6 Tips for Grant Writing

    Type
    Blog Post
    ...in different fields that can provide different insight into an area - not just working within the established...collaborators can also help you get some critical insight into what you're doing wrong. All this helps to...

  9. AAV Production in HEK293 Cells

    Type
    Protocol
    ... through a 0.22 μm membrane. Store at 4 °C. Considerations Before You Start The health of the HEK293T ...vigorously multiple times to obtain a single cell suspension (no clumps of cells). Pool cells from 2 x T-175...forth or shake vigorously to obtain a single cell suspension (no clumps). Count cells using a hemocytometer...above. Add 50 units of benzonase per mL of viral suspension (you should have ~18 mL from steps 12 & 13). ...

  10. Of Myc and Men

    Type
    Blog Post
    ...epitope, a phenomenon known as sequence context sensitivity (Schüchner, et al. 2020). In their study, some... newer Myc-tag antibodies appeared to be less sensitive to sequence context.    So, do Schüchner, et al...

  11. Degrading DNA with Cascade-Cas3

    Type
    Blog Post
    ... increase efficiency of these systems, though consistent expression can still be a challenge. A multitasking...powerful than it is. The nuclease domain of Cas3 is considered weak, especially compared to the power of Cas9...

  12. Tips for a 1st time AAV user (by a Rookie AAV user)

    Type
    Blog Post
    ...primarily found in the culture media, you should consider performing PEG precipitation. Instead of sonicating...solutions are carefully layered and then the viral suspension generated during AAV production is overlaid. ...

  13. 10 Basic tips for mammalian cell culture

    Type
    Blog Post
    ...normally lies around 7.35-7.4 so mammalian cells are sensitive to pH. Anything between orange and light orange...Additional passages will allow your cells to achieve a consistent growth rate. For every minute spent organizing...

  14. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    ..., cultures treated with a folA gRNA were more sensitive to trimethoprim when compared to control gRNA ...for trimethoprim, there would be no changes in sensitivity. Pooled knockdowns are also possible with Mobile-CRISPRi...

  15. 3D Printing Meets CRISPR Cas9

    Type
    Blog Post
    ... 3-D path of the protein’s backbone in three-dimensional space, but their flexible and jiggly nature suggested...structure into experimentally determined electron density, have been known to say something silly like “See...

  16. Antibodies 101: Flow Compensation

    Type
    Blog Post
    ...which way your population needs to be shifted, consider your perpendicular lines. In this case, the population...applying very large compensation values (100+), consider what two markers your fluorescent markers are ...

  17. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    ...nuclease-null—in order to effect different perturbations. Consider, for instance, the challenge of attempting to ...alterations can be exploited to quickly and inexpensively generate a potent Cas9-based transcriptional...

  18. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    ...can survive positive selection, but it will be sensitive to negative selection. Remove the positive selection...creating knockout animals or cell lines, please consider sharing them by depositing with Addgene! Figures...

  20. Light Sheet Fluorescence Microscopy

    Type
    Blog Post
    ... capture large field-of-views (6). For more comprehensive reviews on several other LSFM configurations...Bradke laboratories later developed the 3DISCO (3-dimensional imaging of solvent cleared organs) method of ...

  22. Lentivirus ddPCR Titration

    Type
    Protocol
    ...bleach. Safety Warnings Lentivirus is generally considered biosafety level 2+. Please ensure that you are... media and 11.1 µg/mL Polybrene. Mix the cell suspension well before seeding. Mix each well with a 1 mL...45° angle. Count to 20 while slowly and gently dispensing the droplets down the side of the tube. Place... 95 10 2 1 Denaturation 94 0.5 2 40 Annealing/Extension 60 1 2 40 Enzyme Deactivation 98 10 2 1 Hold 4...

  24. CRISPR 101: RNA Editing with Cas13

    Type
    Blog Post
    ...enzymes do not contain the RuvC and HNH domains responsible for DNA cleavage, so they cannot directly edit.... PspCas13b does not require a PFS, and it is sensitive to mismatches in target RNA from bases 12-26 of...

  26. Course-Based Undergraduate Research Experiences (CUREs)

    Type
    Blog Post
    ...students. But he also found that his teaching responsibilities left him with limited time for one-on-one ...notes the importance of acknowledging student apprehensions about “failure” (protocols not working, hypotheses...

  29. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    ...connections between biology and ancestral groups or, by extension, race. Many researchers do large-scale CRISPR ...potential for bias but to actively look for and consider bias as a confounding factor when interpreting...

  30. Droplet Digital PCR for AAV Quantitation

    Type
    Blog Post
    ...For these reasons, we are in the process of transitioning our titering method from qPCR to ddPCR. Droplet...titer your AAV by ddPCR. Droplet digital PCR is a sensitive assay, so cross contamination into a no template...

  32. Tips for arabidopsis transformation

    Type
    Blog Post
    ...that is wide enough to dunk your plant’s flowers inside. 50 mL falcon tubes or plastic weigh boats are ...from a colony on a plate will reach an optimal density in 18-24 hours. I then add 0.5 mL of this starter...

  33. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    ...non-dividing cells. The first lentiviral vector system consisted of three plasmids: the packaging, envelope, and...capacity of a lentiviral vector but for which transient transgene expression is sufficient. Lentiviral...

  34. Tips for Screening with Yeast Two Hybrid Systems

    Type
    Blog Post
    ...Saccharomyces cerevisiae in 1989 and are still used extensively to screen for molecular interactions in the cell...post-translationally modified, co-expression of the enzyme responsible for the modification in the assay host strain...

  36. Transferable Skills Guide: Creativity

    Type
    Blog Post
    ...with idea generation, try this thought exercise: Consider the issue as if there were no restraints on your...something completely unrelated to the issue and consider how it could be shaped to solve your problem. ...

  37. Hot Plasmids - August 2020

    Type
    Blog Post
    ...CRISPR/Cas Toolkit for plants includes 95 plasmids consisting of CRISPR/Cas nucleases, base editors, gRNA backbones...activation-based norepinephrine (GRABNE) sensors allow for sensitive and high spatiotemporal measurements of norepinephrine...

  38. Plasmids 101: Plasmid Incompatibility

    Type
    Blog Post
    ...partitioning system in the bacteria, but generally it consists of a centromere-like region on the plasmid, a ...2015) Imaging centromere-based incompatibilities: Insights into the mechanism of incompatibility mediated...

  39. How to Be an Excellent Trainee

    Type
    Blog Post
    ...have, then bring them to your mentor. Pro tip! Consider physically walking through the protocol, by going...receive help until you’ve already started a time sensitive experiment. Ready to fly solo? Let your bench...

  40. QC Sequencing Technologies at Addgene

    Type
    Blog Post
    ...with BioRender.com. Sanger sequencing was considered the gold standard until about 2008 when a new...novo assembly within Geneious gives us additional insight into the assembly. Geneious will show how reads...

  41. Twenty Years of Sharing: Addgene's Viral Vector Service

    Type
    Blog Post
    ...our own repository data, giving us significant insight into which vectors are likely to be of most use...Swanson, L. E., & Fan, M. (2016). Practical Considerations for Using Pooled Lentiviral CRISPR Libraries...

  42. Analyzing CRISPR Editing Results with ICE from Synthego

    Type
    Blog Post
    ...editing field by making experimental workflows considerably easier, faster, and more efficient than previous...novice to expert-level researchers in achieving consistently successful CRISPR experiments. Jessica is a ...

  43. Plasmids 101: Stringent Regulation of Replication

    Type
    Blog Post
    ...plasmids tend to be high copy number. Here, we will consider the replication mechanism of plasmids under stringent...plasmid to balance the total plasmid number.  Let’s consider pSC101. This plasmid is stringently controlled...

  44. Mesothelioma - Causes, Symptoms, and Treatment

    Type
    Blog Post
    ...from many other types of cancer. Mesothelioma is considered an orphan cancer; it is rare disease with only...non-resectable mesothelioma by the National Comprehensive Cancer Network, a coalition of the top 27 cancer...

  45. Changing Labor Laws Bring Increased Postdoc Wages

    Type
    Blog Post
    ...technical roles. Therefore, institutions are responsible for ensuring that either all affected scientists...and Brandeis universities have stated that they consider postdocs paid on stipends to be FLSA exempt, and...

  47. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    ...d like to manipulate in mind. Now it’s time to consider where you would like to attack this gene. Should...sequences into HEK293T cells. The fluorescence intensity was classified into four groups (4; brighter than...

  49. Replacing Paper: Tips for Choosing an Electronic lab Notebook

    Type
    Blog Post
    ...this interaction will give you an idea of how responsive and supportive the ELN team is. Once you’ve made...information. If an employee leaves the institution, the transition is smooth as all data is organized within an ...

  50. Five Popular Model Organisms

    Type
    Blog Post
    ...great model organism because they are easy and inexpensive to grow and maintain in the lab, have a short... worm, Caenorhabditis elegans, has been used extensively as a model organism since the 1970s when Sydney...
Showing: 601 - 650 of 881 results