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We narrowed to 869 results for: SOM

Showing: 501 - 520 of 869 results
  1. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    ...knock-ins Are you introducing a large insertion, something 20 base pairs or bigger? Then screening by PCR... pairs modified? In these cases, it’s best to do some planning in the design phase of your experiment ...effects with Sanger sequencing, provided you have some idea of where in the genome you might expect off...
  2. Plasmids 101: Expressing shRNAs for RNA Interference

    Type
    Blog Post
    ...oldie but a goodie — a phrase used to refer to something that has fallen out of fashion but is still useful...either in lentiviral or retroviral backbones. On some occasions, shRNAs are transiently expressed using...strong promoters with no tissue specificity, so in some cases they can lead to toxicity and off target effects...
  3. Antibodies 101: Introduction to Gating in Flow Cytometry

    Type
    Blog Post
    ...Based on personal preference, some people like to make stricter gates while some like to make looser ones ... your cells on a flow plot, you might have to do some testing or ask experienced colleagues.   ...
  4. Targeting HIV-1 with CRISPR: Shock and Kill or Cut it Out?

    Type
    Blog Post
    ...replication. However, this approach could permit some viral protein production, so researchers are also...Cas9 cut site, leading Wang et al. to conclude that some Cas9-derived indels may not ablate viral function...would open up many new therapeutic possibilities for some of the world’s most common diseases. References...
  5. Plasmids 101: Common Lab E. coli Strains

    Type
    Blog Post
    ...different naturally occurring strains of E. coli, some of which are deadly to humans. The majority of all...for growth rpsL Mutation in subunit S12 of 30S ribosome Confers resistance to streptomycin Tn10   Confers...Additionally, Table 2 provides a quick reference for some of the popular strains, their genotypes, and their...
  6. Adenovirus Guide

    Type
    Guide
    ...remain as extrachromosomal elements within the nucleus, normally as double-stranded circular episomes. Due ...restriction enzyme, Gibson Assembly, or Gateway. Some transfer plasmids may have limited restriction sites...be packaged in rAAV vectors. This is the case of some commonly used Cas9 homologs, especially when having...genome and instead are maintained in the cell as episomes. This highly reduces the potential for insertional...nonenveloped viruses from the Adenoviridae family. Episome Genetic element inside the cell that remains as...
  7. Lentiviral Vector Guide

    Type
    Guide
    ...leads to serious disease and occurs in many species. Some common types of lentivirus include Human immunodeficiency...plasmids. Although safer, this system can be more cumbersome to use and result in lower viral titers due to...restriction enzyme, Gibson Assembly, or Gateway. Some transfer plasmids may have limited restriction sites...Standard HEK293T cells are sufficient, but there are some HEK293T cell lines that have been specifically engineered...necessary to properly utilize lentiviral vectors. Some lentiviral plasmids (mostly third-generation) can...Norberg, S. M., Hinrichs, C., Highfill, S. L., Somerville, R. P., Panch, S. R., Jin, P., & Stroncek, D....
  8. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    ... Drosophila or development studies, you may hear some transcription factor and promoter pairs referred...behave very similarly?” The simple answer is that sometimes it is necessary or desirable to control or look...Conclusion This post contains only a brief overview of some of the most common repressible promoter systems,...
  9. Recombinase-based State Machines Enable Order-dependent Logic in vivo

    Type
    Blog Post
    ...enables the design of primer pairs that PCR amplify in some states but not others. Because samples of many registers...output plasmid (single copy, bacterial artificial chromosome backbone) contains the register. Depending on... that, depending on the context and application, some inducible recombinase systems may work faster or...
  10. Generating Mouse Models Using CRISPR/Cas9

    Type
    Blog Post
    ...offspring. Although powerful, this technology is cumbersome to use, not that efficient, often takes more ...must beware that not all gRNAs are created equal. Some work better than others. There are a variety of ...other gRNA design tools available and you can get some additional advice from John Doench’s gRNA design...
  11. PiggyBac-ing Through the Genome Editing Field

    Type
    Blog Post
    ...select, and isolate clones. This procedure is cumbersome and inefficient, as it relies on random integration...there is no selection, you will need to screen somewhere between one hundred and one thousand clones to...boost efficiency over transfecting with plasmids in some cell lines, most likely due to DNA toxicity. More...
  12. Antibody Guide

    Type
    Guide
    ...following table describes the five human isotypes. Some isotypes naturally form multi-antibody units, while...concern for conjugating and signaling molecules, as some buffer components can interfere with the conjugate... Sodium azide, an antimicrobial agent, can block some amines needed for conjugation reactions and should...ubiquitously expressed protein such as GAPDH or tubulin (sometimes referred to as housekeeping genes). This allows...have strong binding between the protein and DNA. Some ChIP protocols use enzyme digestion instead of sonication...antibody using the above process and try again. In some cases, it may be preferable to buy small aliquots...
  13. Promoters

    Type
    Guide
    ...encoding ribosomal RNA (rRNA), which is a main component of a cell’s ribosome structure. Ribosomes are the...and histones (proteins that package DNA into nucleosomes) also bind the TATA box. Histone binding prevents...are responsible for bringing amino acids to the ribosome when proteins are being synthesized. RNA Polymerase...
  14. Plasmids 101: Using Transposons in the Lab

    Type
    Blog Post
    ... themselves from one location and then insert somewhere else (Fig. 2). Class II TEs have LTRs on both ...Applications of transposons Now that you’ve learned about some of the popular transposon systems, let’s take a ...and cell therapy, and have potential to overcome some of the limitations of viruses. TEs have a large ...
  15. Antibodies 101: Antibody Engineering and Directed Evolution

    Type
    Blog Post
    ...goal, and some engineering projects will be easier and quicker than others. We’ve covered some methods ...machinery. But not all engineering is at a large scale — some is done on much smaller equipment on a lab bench...
  16. When Fidelity Matters: A frank discussion about ligase fidelity

    Type
    Blog Post
    ...containing gaps or DNA base pair mismatches (8-11). For some applications, there cannot be any end-joining activity...those containing one or more mismatches. However, some ligases can ligate mismatches to a significant degree...pyrimidine:purine mismatches. Helix stability also plays some role, and mismatches with more hydrogen bonds are...
  17. Fluorescent Proteins 101: Monitoring Cell Mobility Using Fluorescent Proteins

    Type
    Blog Post
    ...swarming around S. aureus and eating the bacteria. Some of the many questions scientists have been able ...used in your experiments, experience has shown that some are more reliable and more efficient at tracking... Tips on Choosing Your Fluorescent Protein Learn Some of the Pitfalls of Working With Fluorescent Proteins...
  18. Cancer and the Immune System: Deciphering the Relationship

    Type
    Blog Post
    ...therapies themselves can be made more effective. Some important factors researchers are beginning to focus...broadly activate the immune response against tumors. Some chemotherapeutic agents induce immunogenic tumor...deaths that help activate tumor responses. Similarly, some radiotherapies have caused increased death receptor...
  19. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...approaches are commonly used to produce KO cell lines, some of which are commercially available. Various proteomic...multiple techniques. For example, we have found that some antibodies perform well in a PFA-saponin fixation...). The approach maps onto the five pillars, with some specific recommendations for how to use these in...
  20. CRISPR/Cas9 FAQs Answered!

    Type
    Blog Post
    ...–4 bp upstream of the PAM sequence. There can be some off-target DSBs using wildtype Cas9. The degree ...efficient guide (most guides would work, but they sometimes have different efficiency). And it is best if ...polymerase or Kapa Hifi Polymerase work very well. Some groups have successfully used Herc II or Kapa Hifi...
Showing: 501 - 520 of 869 results