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We narrowed to 606 results for: abo.2

Showing: 121 - 160 of 606 results

  1. Easi-CRISPR: Generating Knock-In and Conditional Mouse Models

    Type
    Blog Post
    ...process of conventional gene targeting is lowered to about 2-3 months. But certain models remain more difficult...included in the final ssDNA template. Figure 2: Overview of ivTRT to generate an ssDNA repair template... floxed conditional allele requires you to target 2 loxP sites to intronic regions 0.5 - 0.8 kb apart....Typically, Easi-CRISPR works best with insertions of 2 kb or less. Luckily, most of the commonly used cassettes... induction systems (ttA or rtTA) are between 1 to 2 kb in length. There are plenty of Addgene plasmid ...create the ssDNA construct as described in Figure 2.  You can also use Addgene plasmids to purify Cas9... on CRISPR Mouse Models Learn about Homology Directed Repair Learn about Cas9 Ribonucleoprotein (RNP) ...

  2. Optogenetics Guide

    Type
    Guide
    ...influx into the cell when stimulated. Channelrhodopsin-2 (ChR2), the first widely adopted optogenetic tool,... (Phy) interaction 660 Cryptochromes Cryptochrome 2 and CIB1 Blue-light–mediated induction of protein ...interactions based on Arabidopsis thaliana cryptochrome 2 and CIB1 450 LARIAT Inhibits protein function by reversibly...customizable TALE DNA-binding domain with cryptochrome 2 protein and its interacting partner CIB1 to control...transcription effectors OPTOSTIM PHR domain2 of cryptochrome 2 (Cry2) from Arabidopsis thaliana is fused to truncated... with single-spike temporal resolution. PLoS One. 2(3):e299. PMID 17375185 Kim B, Lin MZ. 2013. Optobiology...comparative analysis of microbial opsins. Nat Methods. 9(2):159-72. PMID 22179551 Saunders A, Johnson CA, Sabatini...

  3. Hot Plasmids Podcast Episode 2: New RFPs, AAVs, & More

    Type
    Blog Post
    ...Listen to Our Segement on the Tol 2 Gateway-Compatible Toolbox  Hot Plasmid 2: AAV Vectors for tageting GABA-ergic...of our Hot Plasmids podcast series, you'll learn about new red fluorescent proteins, AAV tools for targeting...

  4. Tips for Getting a Faculty Position

    Type
    Blog Post
    ... this problem? (2-3 sentences) What will be the focus of your first grant? (1-2 sentences) ...of scientific success is publications and grants. 2. Write succinct and easy to understand documents A...consideration. 3. Present an easy to follow seminar See item 2. Attendees will include grad students (that are solicited...people would agree is important to understand. (1-2 sentences). For example: Distinguishing correctly ... potential colleagues study. Think about how you might collaborate or, at the absolute minimum, can appreciate...the first sentence or two what your research is about and what you think you can accomplish. No matter...your seminar accessible. Let the experts ask you about the details and then you can display your deep knowledge...

  5. Retrograde AAV viral preps

    Type
    Collection
    ...Calcium sensor Ting 83899 pAAV-mDlx-GCaMP6f-Fishell-2 mDlx GCaMP6f expression under the control of the mDlx...serotype from the Alla Karpova and David Schaffer laboratories. These viral vector preparations were produced...Janelia Research Campus viral project team in collaboration with David Schaffer (University of California...

  6. 9 Tips to Achieve Success in Academia

    Type
    Blog Post
    ...favorite journals. “If you don't read, you're doomed.” 2. Always be sure to include various positive and negative...will be periods when opportunities arise. “Grab it!” 2. Don’t get caught up in the hype. 3. “Passion gives...sustained motivation is a huge component of success.” 2. “Enjoy Teamwork.” Be friendly, open, sharing, rather... passionate about. Check out my previous post if you're looking for more information about the benefits...every spurious result under the sun. Academia is about convincing results, and nothing is more convincing...is your passion? Even if no one else is excited about it, distinguish yourself with this passion. “Love...presentations and of course, networking and collaborating.” Getting out of the lab also “opens the student's...

  7. 22 Hot Plasmid Technologies from 2014

    Type
    Blog Post
    ...Currently available from the Kim lab are 2 presynaptic and 2 postsynaptic targeting mGRASP plasmids. Additionally... cells were pulsed with 2µg/ml of Doxycycline for 2 hours, and imaged on the Keyence BZ-9000 fluorescence...work! Robinson et al., Dev Cell 2010 Feb 16;18(2):324-31. Cheeseman et al., J Cell Sci 2013 May 1...promotes the formation of an isopeptide bond between 2 peptide tags, SpyTag and KTag. The group demonstrated... of three sets of cloning vectors (Level 0, 1, or 2) which can be utilized in three successive assembly...1 modules can be similarly assembled into a Level 2 vector, thus forming a functional genetic circuit....circuit. Flexibility has been built into the Level 2 vectors to allow for additional iterations of Level 1 assembly...

  8. CRISPR-based Genome Editing Systems For Editing in E. coli

    Type
    Blog Post
    ...plasmids are cured from the cell. Steps 1-1 and 1-2 can be done simultaneously. Image from Li et al., ...1) replaced the promoter of gRNA-pMB1 with PrhaB, 2) changed the replicon of pCas into a non-temperature-sensitive... pEcCas/pEcgRNA system in 4 E. coli K-12 strains, 2 E. coli B strains, 1 E. coli W strain, and 1 Tatumella...MUCICAT better, it is faster. Using MUCICAT only needs 2 newly constructed plasmids (pDonor and pQCascade) ...rounds of transformation in 30 days. Figure 2: (A) MUCICAT plasmids for multicopy integration. (...complete editing process. The MUCICAT system While the above systems are great for experiments that require only...serve to interrupt genes (or gene clusters) in metabolic engineering. So far, we’ve used the MUCICAT system...

  9. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

    Type
    Blog Post
    ...purify IVT sgRNA in the lab. Figure 2: Cas9 RNP complexed with 2-part cr:tracrRNA tags genes more effectively...into stem cells. Cells are allowed to recover for 2-5 days and then analyzed for tagging efficiency using...IVT sgRNA) approach (Dewari et al. 2018 and Figure 2). Not only is the cr:tracrRNA approach highly efficient...cell suspension, transfect and let cells recover for 2-5 days.Note: We use the Nucleofection method for delivery...success was ~%50. If you get low knock-in efficiency (<2% positive by ICC for the tag) or do not see any cells...PMID: 28096221. PubMed Central PMCID: PMC5312033. 2. Dewari PS, Southgate B, Mccarten K, Monogarov G, ...ChIP-seq methods provide a wealth of information about how protein networks work in cells. A major bottleneck...

  10. Plasmids 101: Modular Cloning Applications and Kits

    Type
    Blog Post
    ... Multiple Introns.” Plant Communications, vol. 2, no. 2, Elsevier BV, Mar. 2021, p. 100135. Crossref, ...Constructs.” PLoS ONE, edited by Jean Peccoud, vol. 6, no. 2, Public Library of Science (PLoS), 18 Feb. 2011, p... vol. 5, no. 10, American Chemical Society (ACS), 2 May 2016, pp. 1059–69. Crossref, https://doi.org/10.1021...of Plant Genomes.” The Plant Genome, vol. 16, no. 2, Wiley, 9 Mar. 2023. Crossref, https://doi.org/10.1002...Crossref, https://doi.org/10.1186/s12870-020-02388-2. Stuttmann, Johannes, et al. “Highly Efficient Multiplex...Crossref, https://doi.org/10.1038/s41598-019-46171-2. Resources on Addgene.org Modular Cloning (MoClo... constructs that compose genetic circuits and metabolic pathways. For many MoClo systems, the individual...

  11. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    ..., AAV2/8 has a type 2 ITR and a type 8 serotype. Almost all vectors contain type 2 ITRs, which comes from...differentiated, non-dividing cells.   Figure 2: DNA rescue and transduction of a conventional single-stranded...quantitative PCR-based method for the titration of AAV-2 vector stocks."Molecular Therapy 6.2 (2002): 272. ...make AAV ideal for human therapies and in vivo laboratory experiments ranging from transient gene expression...genetic cargo. Typically, when researchers speak about AAVs, they are actually referring to rAAVs. ITRs...Primer for Beginners Read Our Lentivirus FAQ Learn about Viral Vector Elements Resources on Addgene.org ...

  12. Plasmids 101: FLEx Vectors

    Type
    Blog Post
    ... 2003 PubMed PMID: 12665802. 2. Branda, CS and Dymecki, SM. Talking about a Revolution: The Impact of ...conditional knock-in cassette. As shown in Figure 2, the cassette should contain a reporter DNA sequence...a targeted neuronal population.    Figure 2 (modified from Schnutgen et al. Nat Biotech 2003):... SM. 2008. A FLEX switch targets Channelrhodopsin-2 to multiple cell types for imaging and long-range ... Blog  Read about Other Means of Genome Engineering Using FLEx AAV Vectors? Learn about Viral Vector ...

  13. Controlling for Off-target Effects with a New Genome-wide CRISPR Screen Design

    Type
    Blog Post
    ...guide expression and dsDNA breaks. Figure 2: Non-targeting vs. safe-targeting guides. The genome-wide...target all protein-coding genes present in the genome; 2) non-targeting guides (red), which target no sequence...guides that had zero mismatch off-targets. Guides with 2-bp mismatch off-targets were only toxic if they had...tolerated than mismatches more distal to the PAM, and 2) guides with high GC content have greater off-target...PMID: 28474669. Pubmed Central PMCID: PMC5424143. 2. Tsai, Shengdar Q., et al. "GUIDE-seq enables genome-wide...following unanswered questions about CRISPR screen design. Unanswered questions about genome-wide CRISPR/Cas9...Morgens et al drew some generalizable conclusions about off-target activity, demonstrate safe-targeting ...

  14. Grad School Advice Part 2: Building Community

    Type
    Blog Post
    ...these topics. Listen to Part 1 Here Listen to Part 2 here! Time : Topic 0:00 - 0:50 : Intro 0:51 - 3:08...How did you hear about post grad opportunities and when did you start thinking about them? 18:30 - 19:... top if you leverage your community, think hard about picking a good mentor, and begin considering career...19:54 : How to think about picking a post-doc project 19:55 - 22:40 : Improvements that can be made to ...

  15. How-to: 5 Steps to a Great Panel Discussion

    Type
    Blog Post
    ... let the panelists introduce themselves. I do a 1-2 minute introduction. If you chose good panelists, ...take 5 minutes to introduce themselves and, voila, 1/2 of your time is gone with no interesting discussion...introductions. The moderator asks questions, calling on 1-2 panelist for each question in a way that elicits those...shift, they are checking out. If you can, plant 1-2 good questions with trusted audience members. A good... are nice if you can afford them). I am talking about making sure the audience is interested, educated...of the stories or topics you know they can speak about. There should be some spontaneity, but having a ...location and parking information and remind them about the event as it approaches. Room set-up and logistics...

  16. A Needle in a Base-Stack: Cas9 Structural Biology

    Type
    Blog Post
    ...Cas9 from Streptococcus pyogenes.   Figure 2:  Crystal structure of S. pyogenes Cas9 in the apo...NUC and REC lobes laying open like a book (Figure 2). The portion of the CTD that recognizes the PAM is...it doesn’t appear in the crystal structure (Figure 2) at all. When this crucial recognition region is disordered...regions, so crystal structures like those in Figures 2 and 3 don’t show these regions, even if they are present...itself at the bottom of the image. Stem loops 1 and 2 extend horizontally across both lobes. The guide RNA...is required for Cas9 nuclease function. Stem loops 2 and 3 are not strictly necessary for in vitro cleavage...open conformation, but highly flexible, as in Figure 2. The guide RNA is bound primarily by the REC lobe,...

  17. Addgene is Expanding Our Viral Vector Service!

    Type
    Blog Post
    ...Request review: 1–2 days Prep and QC completed: 6–8 weeks Average shipping time: 1–2 days (USA); 3–4 days... our production process. This process will take 1–2 days, and we anticipate approving the vast majority...educational resources.  We hope you’re as excited about this expansion as we are! Happy packaging! ...

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

    Type
    Blog Post
    ...detect your targeting vector or gene of interest. 2. Breed your parental mice to generate F1 Once you’...homozygous alleles at both of your genes of interest. About 1 in 4 mice will be homozygous for the mutation ...Find our Plasmids 101 blog posts Read blog posts about using Cre-lox Read our genome engineering blog posts... posts Resources on Addgene.org Find more about Cre-lox Read our Genome Engineering Guide Learn more...

  19. An Introduction to Adenovirus

    Type
    Blog Post
    ...either SARS-CoV-2, Adenovirus type 41 (AdV-41), or Adenovirus-Associated Virus type 2 (AAV2) – or some... was either 1) a physiological effect of SARS-CoV-2 toward destabilizing one’s natural immune response...response to Adenovirus (and/or to its partner, AAV) or 2) a sociological effect of the COVID-19 lockdowns causing...… Chiu, C. Y. (2023). Adeno-associated virus type 2 in US children with acute severe hepatitis. Nature...this introduction gives a sense of what AdVs are about – and we hope that it inspires further use of existing...

  20. Rett Syndrome: A History of Research and Therapeutic Outlooks

    Type
    Blog Post
    ...//doi.org/10.1002/1096-8628(200022)97:2<147::aid-ajmg6>3.0.co;2-o Baker SA, Chen L, Wilkins AD, Yu P, ...Caused by a mutation in the methyl-CpG binding protein 2 (MECP2) gene, Rett Syndrome is a rare, progressive...X-linked MECP2, encoding methyl-CpG-binding protein 2. Nat Genet 23:185–188 . https://doi.org/10.1038/13810... (2000) Rett syndrome: Methyl-CpG-binding protein 2 mutations and phenotype-genotype correlations. Am ...therapy. In 2014, RSRT brought together a strong collaboration between two gene therapy labs (Brian Kaspar ...

  21. CRISPR-mediated Plant Base Editors

    Type
    Blog Post
    ...satisfactorily accepts DNA as its substrate (see figure 2).          The artificially evolved adenosine deaminase...editing platforms for plant systems.  Figure 2: Plant base editor platforms and their molecular components...available for plant systems. Target-AID based (CBE-2)-  and rAPOBEC based (CBE-1)-  CBEs are successful...based CBEs are known to deaminate cytosine placed in 2-6 bp and 4-8 bp, respectively, of the protospacer....PMID: 27096365. PubMed Central PMCID: PMC4873371.  2. Nishida, Keiji, et al. "Targeted nucleotide editing...gene, Xw, except for a single base. If you heard about CRISPR, one question probably comes to mind: can...compatibility All of the base editors mentioned above are SpCas9 nickase-based and depend on the presence...

  22. Plasmids 101: Restriction Cloning

    Type
    Blog Post
    ...equivalent to backbone + insert, unsuccessful 2 and 3 above), you can try ligating the recipient plasmid... used. Follow the manufacturer’s instructions.   2. Isolate Your Insert and Vector by Gel Purification...DNA concentration alone. One method is to conduct 2 ligations for each plasmid you are trying to create...containing the ligation mixture with backbone alone; 2. Transformation containing ligation mixture with insert...colonies than the vector + insert plate (unsuccessful 2), or roughly equivalent numbers of colonies on each... (post or pre run in the table above, respectively). Some of the above stains require you to visualize...get experimenting!   Additional Resources: Read about Gel Stains on the BitesizeBio Blog  Additional ...

  23. Viral Vectors 101: What is a Virus (and a Viral Vector)?

    Type
    Blog Post
    ...developed around how they replicate (Figure 2). Figure 2: Baltimore replication classes. Image credit...safer viral vectors that can be used in BSL-1 or BSL-2 labs. Viral vectors are viruses that have been genetically... using biorender.com   If you’re new to talking about viruses, here’s a quick tip on terminology. “Virus...have a growing number of Viral Vectors 101 posts about each type, so be sure to check our blog, or our ...

  24. 15 Hot Plasmids from 2017

    Type
    Blog Post
    ...NGG)  Editing window ~2 nt  85175  pBK-EE-BE3  SpCas9 (NGG)  Editing window ~2 nt  85176  pBK-YE2-...(see figure above). 3 Reasons The PSU Ladders Are Awesome Tools: They are comprised of 2 plasmids, pPSU1...quantum yield of 0.54) and mScarlet-H with M164H (2-fold improvement in photostability compared to mScarlet...SpCas9 base editors with editing windows as small as 1-2 nucleotides! To reduce off-target effects associated...pBK-YE2-BE3  SpCas9 (NGG)  Editing window ~2 nt  85177  pBK-YEE-BE3  SpCas9 (NGG)  Lower on-target activity...activity than BE3; editing window ~1-2 nt  87438  pET42b-HF-BE3  HF-Cas9 (NGG)  Very low off-target activity...libraries The Gross Lab at UCSF recently deposited 2 Bacillus subtilis libraries with Addgene – the B. ...

  25. Viral Vectors 101: AAV Variables That Matter

    Type
    Blog Post
    ...testing your AAV.  Figure 2: Comparison of AAV serotypes 1, 2, 5, 6, 8, and 9 in two different .... Factors to think about are (1) your experimental conditions and measurements; (2) the cells you’re targeting... Tropism and Axonal Transport of AAV Serotypes 1, 2, 5, 6, 8 and 9 in the Mouse Brain. PLOS ONE, 8(9),... Comparison of four promoters. Gene Therapy, 28(1–2), 56–74. https://doi.org/10.1038/s41434-020-0169-1...analyses of adeno-associated viral vector serotypes 1, 2, 5, 8 and 9 in marmoset, mouse and macaque cerebral...

  26. Hot Plasmids and Viral Preps - January 2021

    Type
    Blog Post
    ... Opto-Cre-Vvd is fast and can cut in as little as 2 hours.  Sheets, MB, et al., ACS Synth Biol. 2020....they investigated three Casɸ orthologs, Casɸ-1, Casɸ-2, and Casɸ-3. This system is notable for several reasons...a minimal PAM requirement. For instance, for Casɸ-2 the PAM sequence is 5’-TBN-3’ (where B is G, T, or...deposited by Leopold Parts. This library delivers 2 randomly paired guides per construct and allows a ... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here.  Here's...

  27. Using AAV for Neuronal Tracing

    Type
    Blog Post
    ...for retrograde transport has been found for AAV1, -2, -5, -7, -8 (Taymans et al., 2007), AAV1 (Hollis et... tropism and axonal transport of AAV serotypes 1, 2, 5, 6, 8 and 9 in the mouse brain. PLoS One 8:e76310...reveal novel hippocampal CA2 circuits. Nat Neurosci 17(2), 269–279. PubMed PMID: 24336151. PubMed Central PMCID... Mechanisms of Motility and Regulation. Neuron 84(2), 292–309. PubMed PMID: 25374356. PubMed Central PMCID...transduction efficiency of recombinant AAV serotypes 1, 2, 5 and 8 in the rat nigrostriatal system. J Neurochem...analysis of adeno-associated viral vector serotypes 1, 2, 5, 7 and 8 in mouse brain. Hum Gene Ther 18, 195–...retrograde access to projection neurons. Neuron 92(2), 372–382. PubMed PMID: 27720486. PubMed Central PMCID...

  28. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    ...the absence of genome editing (Fig. 2 and 3) (5, 3). Figure 2: Gene activation with nuclease-competent...shown to exhibit more limited activity than SpCas9 (2). In many cases this is due both to lower relative...PMID: 24076762. PubMed Central PMCID: PMC3844869. 2. Chari, Raj, et al. "Unraveling CRISPR-Cas9 genome...of the Addgene Scientific Advisory Board. His laboratory at Columbia University is focused advancing scientific... Listen to Our Interview with Alex Chavez Learn about Sequencing Options for CRISPR Genotyping Cas9 Activators...

  29. Year of the Camelid: Antibody Style

    Type
    Blog Post
    ...and a source of sdAbs (Figure 2) (Xu, et al., 2021).      Figure 2: Genetically modified mice offer... from camelid mice and llamas neutralize SARS-CoV-2 variants. Nature 595:278–282. https://doi.org/10.1038...the commercial "Nanobody" name.  We’ve talked about sdAbs a number of times on this blog over the years...

  30. Zhang Lab's CRISPR Frequently Asked Questions

    Type
    Collection
    ...extraction solution, runs the protocol, then uses around 2-4ul in each 50ul PCR reaction. The gDNA extracted ... Feng Zhang's Lab, has assembled a list of FAQs about using the lab's CRISPR technology. Genome..., February 2013 ), has assembled a list of FAQs about using the lab's CRISPR technology. Visit the Zhang...

  31. Antibodies 101: Chimeric Antibodies

    Type
    Blog Post
    ...use 2 different secondaries for your antibody pair and visualize both targets in parallel (Fig. 2). For...successfully visualize both proteins.     Figure 2: Two primary antibodies with the same isotype can ...flexibility needed for multiplexing. In the case above, you can’t costain with your desired antibodies ...multiplexing with chimeric antibodies. In the case above you should use an anti-rat IgG1 secondary antibody...this blog post has answered all of your questions about the utility of isotype conversions and chimeric ..., Bright, S. M., Robinson, M. K., Graham, E. T., Sabo, J. P., Owens, R., King, D. J., Norris, S. H., &...

  32. Tetbow: Bright Multicolor Labeling for Neuronal Tracing

    Type
    Blog Post
    ...possible when each of the XFP genes is introduced at ~2 copies per cell per color, following a Poisson distribution...(prepared with Addgene #104110 - #104112) (Figure 2). We’ve also created constructs with containing chemical...AAV titer - The XFP genes have to be introduced at ~2 copies per cell per color to achieve the highest color...Additional resources on the Addgene blog Read about Brainbow Learn about using AAVs for neuronal tracing Visit...page Additional resources on Addgene.org Learn about Addgene's viral service See our fluorescent protein...

  33. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    ...procedure to be performed using a simple two-step, 2 hr protocol prior to transformation (Figure 1). As...Primer design Using specific primer design (Figure 2), IVA cloning can be used to perform any plasmid modification... for each type of modification is shown in Figure 2, and is discussed in more detail below. The main requirement...Biotechniques 10.1 (1991): 62-66. PubMed PMID: 2003926. 2. Bubeck, Peter, Monica Winkler, and Wilfried Bautsch... Watson and Javier García-Nafría from the MRC Laboratory of Molecular Biology. Plasmid cloning is an essential...with successful recombination reported in all laboratory E. coli strains tested so far. The use of this...low cost and can be immediately adopted by any laboratory. All procedures are performed using the same...

  34. Fluorescent Proteins 101: Aptamer Fluorophores

    Type
    Blog Post
    ...Fig.2). Spinach eventually evolved into Spinach2, a more stable and brighter version of the aptamer, but...PMID: 25713381. PubMed Central PMCID: PMC4364232. 2. Baker, Monya. “RNA imaging in situ”. Nature Methods...emerging environmental pollutants.” Front Chem. 2014; 2: 41. PubMed PMID: 25019067. PubMed Central PMCID: ...biosensors.” TrAC Trends in Analytical Chemistry 27(2):108-117 · February 2008. 11. Svensen, Nina and Jaffrey...target metabolite creates an aptamer that will only bind to DFHBI in the presence of that metabolite (Fig...approach less sensitive in most applications. Learn About Fluorescent Biosensors Light-up aptamers (or, fluorescing...fluorophore protein. Years later, Samie Jaffrey’s laboratory took advantage of their deep knowledge of the...

  35. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    ...the light-inducible heterodimerizing cryptochrome 2 (CRY2) and calcium and integrin-binding protein 1 ...inactive, dead Cas9 protein (dCas9) fused to CIB1; and 2) the activator - the CRY2 photolyase homology region... fused to both the N- and C-termini of dCas9; and 2) CRY2FL-VP64, a fusion of full-length CRY2 and the...researchers saw an 11-fold increase in mRNA levels after 2 hr and a 400-fold increase after 30 hr. The system...authors fragmented Cas9 into N-terminal (residues 2-713, N713) and C-terminal (residues 714-1368, C714...nMagC714 instead of nMagHigh1-C714, generating paCas9-2. This change did not significantly alter the system...prior work, the Sato lab also showed that the paCas9-2 system could be spatially controlled and reversibly...

  36. Delivery Methods for Generating iPSCs

    Type
    Blog Post
    ...plasmid-based expression requires serial transfection of 1 or 2 plasmids that express the reprogramming factors of...for generating iPSCs by plasmid transfection where 2 of 11 clones tested had plasmid integration. Another...understanding the mechanisms of reprogramming, and 2) studies with clinical endpoints. In the first scenario...PubMed PMID: 27764162. PubMed Central PMCID: 5072705. 2. Sommer, Cesar A., and Gustavo Mostoslavsky. "The ...challenges." Journal of Cellular Physiology 228, no. 2 (2012): 267-75. PubMed PMID: 22767332.  3. Kim, Dohoon...PubMed PMID: 23546745. PubMed Central PMCID: 4176696. 2. Yusa, Kosuke, Roland Rad, Junji Takeda, and Allan...generation. Expert opinion on biological therapy, 10 2, 231-42. PubMed PMID: 20088717. PubMed Central PMCID...

  37. Addgene’s Newest Way to Speed Your Research: The Cloning Grade DNA Service

    Type
    Blog Post
    ...plasmid tools. Details about the cloning grade DNA beta project We will ship you  2 µg of DNA in 20 µl of... be directly used in the research on the SARS-CoV-2. Nine of the overall plasmid selection will be open...Plasmids 101 blog series Learn about the different grades of DNA  Read about the various ways to quantify...quantify DNA Resources on Addgene.org Learn about our other beta projects Read our Molecular Biology Reference...

  38. CRISPR 101: Making a Knock-In Cell Line

    Type
    Blog Post
    ... However, if you have a larger knock-in (200 bp – 2 kb), a dsDNA donor will need to be used due to synthesis...accidentally change your protein of interest.   Fig. 2: Stop re-cutting with PAM and guide RNA disrupting... the other repair mechanisms Mammalian cells have 2 primary repair mechanisms besides HR - Non-homologous.... Com., 8(2024), (2017). 10.1038/s41467-017-01836-2 Charpentier, M., Khedher, A. H. Y., Menerot, S., Brion...CRISPR Featured Topic Page Learn about Non-Homologous End Joining Learn about Base Editing Read 3 Tips to ... post on increasing HDR efficiency to read more about optimizing this distance. Homology arms and types...screening strategies for your experiment. Also learn about Addgene’s knock-in plasmids for reagents to help...

  39. Lentiviral Vector Uses and Overview

    Type
    Blog Post
    ...create the 2nd-generation lentiviral system (Figure 2). The HIV genes that do remain are very important ...1996 Apr 12;272(5259):263-7. PubMed PMID: 8602510. 2. A third-generation lentivirus vector with a conditional...CH, Humeau LM, Rebello T. J Gene Med. 2013 Feb;15(2):78-82. PubMed PMID: 23322669. 6. Genetic treatment... Banasik MB, McCray PB Jr. Gene Ther. 2010 Feb;17(2):150-7. PubMed PMID: 19847206. Additional Resources...gene therapy applications. Read on to learn more about the current (and future) applications of lentiviral...do both, Addgene depositor Didier Trono and collaborators turned to the lentivirus HIV-1, well-known to...Libraries Enable Genome-Scale, Knockout Screening Read about Genome-Wide Screening Using CRISPR/Cas9 Additional...

  40. Deep Dive: Statistical Tests (Comparisons)

    Type
    Blog Post
    ...typically, normal (bell curve) or skewed (see Fig. 2). Assumptions – characteristics of a data set; ...levels of variance among data.     Fig. 2: Examples of data distribution profiles.   Parametric... positive is much higher than if you only look at 2 or 3 pairwise comparisons. Similar to family-wise ...statistical facets of reproducibility. Adv Physiol Educ. 40(2): 248-252 (2016). 0.1152/advan.00042.2016 Finch, W...Simulation Study. Front. Appl. Math. Stat. Volume 2 (2016). doi.org/10.3389/fams.2016.00002  ...a summary snapshot without telling you anything about outliers or data spread within those data sets. ...the rules” and which we have certain information about. The assumptions and requirements that need to be...
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