Skip to main content
Addgene

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

In Category

We narrowed to 297 results for: hal.2

Showing: 161 - 180 of 297 results

  1. Using Video to Share Your Science: We Share Science

    Type
    Blog Post
    ...Information Science and Technology 60.4 (2009): 858-862. 2. Van Noorden, Richard. "Interdisciplinary research...Librarianship and Scholarly Communication 2.2 (2014): 2. Additional Resources on the Addgene Blog 6 Tools...solutions to complex challenges. As a cross-disciplinary researcher, the challenge of keeping up on studies... Sharing your research with the world can be challenging. After months, or years, of grueling effort to...biology or international relations.  Many of the challenges in science today are too complex for any single...making publishing their research all the more challenging even when it is of quality). On the site you ...

  2. A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells

    Type
    Blog Post
    ...NGG-3′ (“N” can be any nucleotide base, followed by 2 guanines). This limits the use of ABEs as DNA sites... in the promoter region of gamma hemoglobin (HGB1/2) leads to continuous expression of fetal hemoglobin...T-to-G•C base changes (Figure 1), but it can be challenging to make these base changes, especially in primary...𝛽-globin), which cause sickle cell disease and 𝛽-thalassemia, can be mitigated with the expression of fetal...

  3. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    ... digest and compare it to some reliable controls. 2. Viral stocks may be sensitive to freeze thaw cycles...PMID: 24003179. PubMed Central PMCID: PMC3760253. 2. Rohwer, Forest. "Global phage diversity." Cell 113.2...tasks, both of which are factors in the technical challenges they can present. Many researchers find themselves...after transfecting cells, replace the medium with half the volume typically used (e.g., use 5 mL medium...

  4. Deep Dive: Fixing and Permeabilizing for Immunofluorescence

    Type
    Blog Post
    ... of permeabilization and membrane integrity (Fig. 2). For example, saponin interacts with cholesterol ... at too high a concentration.      Figure 2: Examples of different permeabilization strategies...samples in a given state or point in time. You want to halt degradative processes while maintaining the structure...fixation-artifacts-and-how-to-minimize-them/  Goldenthal KL, Hedman K, Chen JW, et al (1985) Postfixation...

  5. FlipGFP, a novel fluorescence protease reporter to study apoptosis

    Type
    Blog Post
    ...parts: (1) β-strands 1-9 (β1-9) and the one ɑ-helix, (2) β-strand 10 (β10), and (3) β-strand 11 (β11). Each...activating caspase-3, and detected fluorescence within 2 to 5 hours, a time consistent with previous methods...fluorescence imaging of living organisms can be challenging due to issues with tissue autofluorescence, cell...

  6. Advice for Starting a Biotech Company from a Grad Student turned Entrepreneur

    Type
    Blog Post
    ...go anywhere. The team should initially consist of 2-3 individuals that are aligned in vision and outcomes... the technology. This transition takes around 1 – 2 years. The product is the measure of success. Technology...his co-founders raised $6 million. The biggest challenge during this time was learning to communicate effectively...

  7. Your Top Requested Plasmid in 2015!

    Type
    Blog Post
    ...PubMed Central PMCID: PMC4089965. 2. Sanjana, Neville E., Ophir Shalem, and Feng Zhang. "Improved vectors...only been available for a little over a year and a half, a small sampling of the research it has already...another great year of sharing!   References 1. Shalem, Ophir, et al. "Genome-scale CRISPR-Cas9 knockout...

  8. Fluorescent Proteins 101: Fluorescent Protein Timers

    Type
    Blog Post
    ...linear region to solve for total expression time. 2. Timer functionality with a single gene Although it...290.5496 (2000): 1585-1588. PubMed PMID: 11090358. 2. Tsuboi, Takashi, et al. "Age-dependent preferential...understand how protein localization changes over time.Hallmarks of fluorescent protein timers When FP timers...noticed a few properties that have now become hallmarks of ideal FP timers: 1. Ratiometric determination...

  9. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...Workflow Timeline Day 0: Seed and transduce Cells Day 2–3 (am): Remove media, replace with fresh media containing...Biological Safety Cabinet 0.5–10 µL single channel pipette 2–20 µL single channel pipette 20–200 µL single channel... 200–1000 µL single channel pipette Ice bucket CO 2 incubator Pipet controller Hazardous waste container...cell death, the cell media should be changed every 2–3 days to maintain the dose of antibiotic, which may...confluent 10 cm dish can be expanded into two 75 cm 2 flasks, etc. Pro-Tip This selection method results...genome-wide libraries for CRISPR screening. Sanjana NE, Shalem O, Zhang F. Nature Methods. 2014 Aug;11(8):783-...

  10. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    ...immediate early gene expression. Genes Dev. 1988 Jun;2(6):718-29. doi: 10.1101/gad.2.6.718. PMID: 2843425...Targeted Gene Expression in Drosophila.” Genetics 186(2) (2010): 735-55. PubMed PMID: 20697123. PubMed Central... in Saccharomyces cerevisiae.” FEMS Yeast Res. 12(2) (2012): 197-214. PubMed PMID: 22129153. Additional...solve the problem of broad QF expression-induced lethality in flies. Riabinina et al. also developed chimeric...

  11. Our Most Popular Blog Posts from 2019

    Type
    Blog Post
    ...fusion location, there are many choices to make. 2. Prime Editing: Adding Precision and Flexibility to...and represent the diversity of tools we share on behalf of scientists. 9. Plasmids 101: Positive and Negative...fluorescent proteins at Addgene. 7. Designing Your Chalk Talk for the Academic Job Interview by Erik Snapp...blogger Erik Snapp breaks down how to prepare a chalk talk for an academic job interview including defining... research vision, organizing your vision into a chalk talk, and tips for practicing.  6. The Fluorescent...

  12. CRISPR Library Amplification

    Type
    Protocol
    ... recover, set up overnight growth (Estimated time 2-3 hours) Transformation should be performed at the... day to ensure that growth times are limited. Day 2: Harvest cells and purify DNA (Estimated time 3-4 ... Tubes should contain a total of 5 mL (3 mL SOC + 2 mL transformed Endura from two separate transformations...has been absorbed by the agar. This usually takes 1-2 minutes. Critical Be careful not to rip or shred the...incubation if needed. Place 100 mL sterile LB at 4 ℃. Day 2 (morning) Before beginning, prechill at least four...pellet. The total weight of each pellet should be ~1-2 g. Pro-Tip Make sure to weigh the empty tube beforehand...Quality Control (QC) Pooled libraries can be challenging and expensive to ensure adequate quality, but...

  13. CRISPR Activators: A Comparison Between dCas9-VP64, SAM, SunTag, VPR, and More!

    Type
    Blog Post
    ...gene activation, with some genes experiencing around 2-fold activation levels. Although other methods have.... This method has been shown to activate genes at 2- to 3-fold higher than SAM, SunTag, and VPR (Zhou ...Ter-Ovanesyan D, Haque SJ, Cecchi RJ, Kowal EJK, Buchthal J, Housden BE, Perrimon N, Collins JJ, Church ...

  14. PCR Applications for Copy Number Variation Assays

    Type
    Blog Post
    ...mean you have one copy of the gene; an RQ value of 2 means you have two copies, and so on and so forth....quantitative PCR. Expert Rev Mol Diagn. 2005 Mar;5(2):209-19. doi: 10.1586/14737159.5.2.209. PMID: 15833050...available, ddPCR may be a better option for more challenging cases.  Price point If budget is a concern...

  15. PITChing MMEJ as an Alternative Route for Gene Editing

    Type
    Blog Post
    ... PITCh vector via In-Fusion or SLIC cloning. Step 2: Design an insertion locus-specific gRNA The gRNA ...Find the plasmids from this publication at Addgene. 2. Dalvai, Mathieu, et al. (2015). “A Scalable Genome-Editing...cell lines can be created in about a month and a half, without the need for complicated cloning of homology...

  16. Chemogenetics Guide

    Type
    Guide
    ...muscarinic receptor and has not been used in vivo . Figure 2: DREADDs, their ligands, and signaling properties.... Reference Rq(R165L) Human M3 muscarinic Arrestin-2/-3 CNO* Increase Arrestin translocation Arrestin signalling...hM1Dq hM5Dq Human M3 muscarinic G αq CNO* Increase Ca 2+ Neuronal burst firing Armbruster et al., 2007 rM3D...channels, their effect, and outcome in neurons. Table 2. Descriptions of PSAMs and their activity in neurons...Forbes SL, Liggett SB (2001). Modification of the beta 2-adrenergic receptor to engineer a receptor-effector...Ikefuama EC, Slaviero AN, Silvagnoli AD, Crespo EL, Schalau R, Gott M, Tree MO, Dunbar GL, Rossignol J, Hochgeschwender...

  17. Fluorescent Proteins 101: Green Fluorescent Protein (GFP)

    Type
    Blog Post
    ...from GFP and the mutation definitions found in Table 2 may not apply. Table 1: The specific mutations comprising..., V163A EBFP F64L, S65T, Y66H, Y145F   Table 2: Functional role of specific mutations in GFP derivatives...and when they come into close proximity, the two halves of EGFP undergo folding, maturation, and fluorescence...

  18. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...components, see the AAV Plasmid Elements table . Figure 2: Plasmids required for rAAV vector production. Created... pseudotyped with viral capsids from serotypes 1, 2, and 5 display differential efficiency and cell tropism...the central nervous system . Molecular Therapy, 10 (2), 302–317. https://doi.org/10.1016/j.ymthe.2004.05.024...the production of AAV vectors . Microorganisms, 12 (2), 384. https://doi.org/10.3390/microorganisms12020384...retrograde access to projection neurons . Neuron, 92 (2), 372–382. https://doi.org/10.1016/j.neuron.2016.09.021...short coding sequences of channelrhodopsins, halorhodopsins, and other optogenetic genes makes them suitable...) and a gRNA together into an rAAV capsid is challenging due to its limited packaging capacity, especially...

  19. Finding Your Science Policy Path

    Type
    Blog Post
    ...teaching and mentoring others (see more in Figure 2). Starting early demonstrates not only your interest...application process so you can learn more! Figure 2: The many ways to prepare for a science policy career...That’s what makes it so wonderful. The most challenging part of jumping into this career path from academia... your hair out, your successes, failures, and challenges you’ve overcome. This information may help you...completely new field without issue. This may be more challenging if you are trying to apply for an outright job...

  20. Better Dyeing Through Chemistry & Small Molecule Fluorophores

    Type
    Blog Post
    ...expressed as a fusion with your favorite protein and (2) a synthetic fluorophore-contianing “ligand” that ...cell 58.4 (2015): 644-659. PubMed PMID: 26000849. 2. Grimm, Jonathan B., et al. "A general method to improve... include: Self-labeling tags (e.g., SNAP-tag, HaloTag, TMP-tag) – These widely used systems consist of...
Showing: 161 - 180 of 297 results