Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of the Addgene plasmid sequence database.

Information

In Category

We narrowed to 730 results for: NSI

Showing: 221 - 240 of 730 results

  1. Flow Cytometry Readouts: Yes, No, and Everything in Between

    Type
    Blog Post
    Published
    Aug. 20, 2024, 1:15 p.m.
    ...positive population is no higher than 105 signal intensity. Signals any higher than this may not be clearly...comparison, say somewhere between a 104–105 signal intensity. If your samples and controls were processed ...better. The histogram shows peaks of high signal densities, so signal shifts due to upregulation or downregulation... shifts in signal using the mean fluorescence intensity (MFI) of the different populations. MFI is the...

  2. Human Germline Editing Using CRISPR

    Type
    Blog Post
    Published
    Aug. 10, 2017, 2:19 p.m.
    ...clinical applications could be considered. It is important to consider the question of broader applicability...disease prevention. However, it is important to consider that already-tested CRISPR alternatives are applicable...many more experiments are needed before we can consider clinical applications for germline editing. Although...

  3. To Each HIS Own

    Type
    Blog Post
    Published
    March 2, 2023, 2:15 p.m.
    ... 2000).  One downside of polyhistidine tag-based purification, however, is the propensity for nonspecific...contamination, such as mammalian production systems, consider using these more selective substrates along with...the opposite end.  If neither placement works, consider purifying under denaturing conditions. Unlike ...

  4. All in a Twist: dsRNA

    Type
    Blog Post
    Published
    Feb. 2, 2023, 2:15 p.m.
    ...process. These transcription-coupled R-loops are transient in nature and will dissociate when gene transcription... respond accordingly. While not perfect, these sensing systems work to distinguish between RNA of cellular...viral origin and respond appropriately. When these sensing systems fail, diseases and steep cellular consequences...or anticipating this response are important considerations when using these reagents. RNA:DNA hybrids...

  5. Supporting Rare Disease Research with the 2020 BeHEARD Awards

    Type
    Blog Post
    Published
    Sept. 8, 2020, 2 p.m.
    ...to a large family of solute channels that are responsible for maintaining the nutrient and ionic balance...says Iyer. Since the RESOLUTE plasmid collection consists of human and murine solute channels, the team ... University.  These protein-receptor complexes consist of many parts that work together on the cell membrane...transduction events, we will utilize BRET-, FRET-, and intensity-based signaling biosensors to monitor intracellular...

  6. Plasmids 101: Golden Gate Cloning

    Type
    Blog Post
    Published
    Aug. 27, 2015, 2:30 p.m.
    ...vector + insert may spontaneously religate, this transient construct retains functional Type IIS sites and...be scarless. Golden Gate assembly is also less expensive than many commercial cloning methods. Golden ...amenable to "domestication", you might want to consider using an alternative method like Gateway cloning...cloning or Gibson assembly. Another important consideration is the design of flanking overhangs. Although...

  7. Working with Nuclear Receptors

    Type
    Blog Post
    Published
    July 15, 2014, 4:03 p.m.
    ... multitude of therapeutic indices, including hypertension, cancer, diabetes, cardiovascular disease, cholesterol...receptor research, and has generated a set of comprehensive datasets that collectively address many important...of coregulators in HeLa cells was the first comprehensive analysis of the interactome of these key regulatory... between laboratories. The development of the transient transfection platform for assaying transcriptional...

  8. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    Published
    Sept. 28, 2020, noon
    ... in E. coli. The system available from Addgene consists of two plasmids: pCas9: carries Cas9 and chloramphenicol... enable gRNA-directed transposition. INTEGRATE consists of four major components including (1) a CRISPR... ability to multiplex repression. Using a Tet responsive promoter to drive expression of a catalytically...and developed new, more tightly controlled, Tet responsive promoters to avoid leaky activation of the system...technology can have on the human microbiome. You can consider the microbiome as a collection of organisms, their...

  9. Hot Plasmids and Viral Preps - March 2021

    Type
    Blog Post
    Published
    March 23, 2021, 1:15 p.m.
    ...categories: probes and editors. A probe array may consist of at least two sensor modules targeting a specific...auxin inducible degron (AID) system has been extensively used in C. elegans since 2015 to conditionally... which can be easily visualized.  The tool-kit consists of highly useful vectors compatible with Gibson.... Check out the new additions.  Image calcium transients in PV interneurons with AAV-S5E2-GCaMP6f, now...

  10. Design Tips for Prime Editing

    Type
    Blog Post
    Published
    Jan. 23, 2025, 2:15 p.m.
    ...a prime editing guide RNA (pegRNA). The pegRNA consists of a CRISPR gRNA extended to include a primer ... BioRender.com. The first base of the 3′ extension of the pegRNA should not be C. A C base is speculated...al., 2014). Figure 3: pegRNAs whose 3′ extension begins with a C base can cause unwanted base pairing...influence the success of your prime edit. If possible, consider following these suggestions: Edit the PAM along...

  11. 7 Simple Strategies to Resolve Conflicts with Difficult Supervisors

    Type
    Blog Post
    Published
    Jan. 21, 2015, 1:30 p.m.
    ...time moaning about their troubles, Silent or unresponsive clams, who seem to ignore you and your questions...communicate their ideas confidently while also considering the needs of other people. Some students and ...for it. Do not take criticism personally or get defensive. Look at the problem from their viewpoint, and... you even when you meet in person (silent or unresponsive clam), you will probably need support from your...

  12. Plasmids 101: Protein Expression

    Type
    Blog Post
    Published
    June 7, 2018, 1:17 p.m.
    ...Overall the general strategy for protein expression consists of transfecting cells with your DNA template of... Mammalian systems can express proteins both transiently or through stable cell lines. Both methods produce...recombinant proteins from plants however can be expensive and time consuming as plant tissues themselves...applications. Cell-free expression systems however, are expensive and very technically challenging to use. Alyssa...

  13. When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells

    Type
    Blog Post
    Published
    April 19, 2016, 2:30 p.m.
    ...all seriousness, EGFP/GFP has a real nontrivial propensity to noncovalently dimerize. That means that your...to not produce artifacts in your experiments. Considerable effort has gone into making FPs monomeric, yet...actually monomeric, at least in practical terms. The propensity of a particular type of molecule to form a dimer...study of GFP-based FRET, with application to intrinsically unstructured proteins. Protein Sci, 2007. 16...

  14. PCR Applications for Copy Number Variation Assays

    Type
    Blog Post
    Published
    April 26, 2022, 1:15 p.m.
    ...most starting materials, and is not particularly sensitive. The fluorescent signal can be affected by AT-...Image courtesy of: Ygonaar    Taqman is more sensitive than Sybr, with less background noise, and is ...amplified in the same reaction. Taqman’s increased sensitivity allows for a more confident determination of ...approach is right for your experiment. You’ll have to consider:  Resources needed Most molecular biology labs...

  15. The AAVantages of AAV in CRISPR Screens

    Type
    Blog Post
    Published
    Feb. 4, 2025, 2:15 p.m.
    ...gotten larger, easier, and more comprehensive thanks to the consistent development and improvement of ... for each experiment. Additional considerations One of the downsides to using AAV is the limited packaging...

  16. PRIDICT: Predicting Efficiencies of Prime Editing Guide RNAs

    Type
    Blog Post
    Published
    June 25, 2024, 1:15 p.m.
    ...The pegRNA consists of a target-complementary spacer, a scaffold sequence, and an extension sequence containing...screens such as PRIDICT/PRIDICT2.0 provide valuable insights into pegRNA editing efficiencies for specific ...experiments, thus saving you time, sweat, and money on extensive optimization efforts. This holds true for any ...

  17. Antibodies 101: Conventional vs Spectral Flow Cytometry

    Type
    Blog Post
    Published
    Aug. 3, 2023, 1:15 p.m.
    ...which spectral flow cytometry requires special considerations. Conventional flow cytometry During conventional...cytometry for a given experiment, it is important to consider how many parameters you want to use in a given...and spectral flow cytometry, it is important to consider the expression levels of the markers you are including...molecules conjugated together) can be especially sensitive to batch effect. For example, when a PE molecule...

  18. Transgenic Organisms, Cas9 Gene Drives, and Appropriate Safeguards

    Type
    Blog Post
    Published
    May 22, 2015, 4:58 p.m.
    ...RNA-guided gene drives in the summer of 2014 (after extensive discussions with biosecurity experts, ecologists...gene drives intended for eventual release should consider building an appropriate immunizing reversal drive...not use them whenever applicable?Against this, consider the cost of an accidental release. Science relies...trusted to handle such a powerful technology responsibly. An accidental release would be particularly ...

  19. Tips for Using BLAST to Verify Plasmids

    Type
    Blog Post
    Published
    May 29, 2014, 1:29 p.m.
    ...DDBJ, and PDB sequences and may be the most comprehensive for searching. Timesaving Tip #1: If you know...searches! Now, before you click the BLAST button, consider the Program Selection parameter, as this will ...the alignment at a more distal amino acid. We consistently run our blastx searches with the “Low complexity...101: How to Verify Your Plasmid Plasmids 101: An Inside Look at NGS Plasmid Quality Control Learn about...

  20. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    Published
    July 31, 2018, 12:55 p.m.
    ...abandoned early on due to its considerable immunogenicity and its transient expression. LV results in long...been shown to to transduce the retina but only transiently. However new viral vectors capable of providing...disease-causing genes, including the RPE65 gene responsible for an autosomal recessive, severe, childhood-onset...
Showing: 221 - 240 of 730 results