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We narrowed to 608 results for: Sis

Showing: 161 - 200 of 608 results

  1. Interview: Nicola Patron on Plant Synthetic Biology, MoClo, and More

    Type
    Blog Post
    Published
    June 12, 2014, 3:30 p.m.
    ...plants, and agricultural crops in particular, resistant to disease. They also devise biosensors designed... developed mostly with model plants in mind, Arabidopsis and Nicotiana benthamiana. N. benthamiana is ...Universities sign up and in the first year there is an analysis of women in faculty, how they are doing. They ...Plant SynBio Reading: Quick, Versatile Plant Transgenesis with GreenGate Plasmids  ...

  2. Tips for Using BLAST to Verify Plasmids

    Type
    Blog Post
    Published
    May 29, 2014, 1:29 p.m.
    ...the alignment at a more distal amino acid. We consistently run our blastx searches with the “Low complexity...recommendation is not infallible, we have found it saves analysis time to remove this default setting. Timesaving...Depending on your sequencing result and desired analysis, BLAST may not always be your optimal choice. ...plasmid maps. Resources on Addgene.org Sequence Analysis of your Addgene Plasmid Addgene Plasmid Guide ...

  3. Introducing Addgene's New Search - Find Plasmids More Easily For Your Research!

    Type
    Blog Post
    Published
    Dec. 3, 2019, 11:38 a.m.
    ...bacterial resistance marker used to grow plasmid containing bacterial cultures Bacterial Resistance: Use this...terms with gene and guide RNA / shRNA inserts Resistance Marker: This indicates the selectable marker ... this to choose the antibiotic resistance gene(s) used for selection in bacteria. Has Viral Service?: ...

  4. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    Published
    July 31, 2018, 12:55 p.m.
    ... rare genetic eye disease, Leber’s congenital amaurosis. This is the first of what I, scientists, doctors...disease more prevalent or life-threatening (cystic fibrosis, hemophilia, muscular dystrophy)? AAV gene therapy...childhood-onset form of blindness name Leber’s Congenital Amaurosis (LCA). Shortly after the cloning of the RPE65 ... time and place. But it is the hard work and persistence of a dedicated team of scientists led by Dr. ...

  5. Isolating Genomic Regions of Interest with the CRISPR System

    Type
    Blog Post
    Published
    May 30, 2019, 12:44 p.m.
    ...specific genomic region using the CRISPR system, consisting of dCas9 and targeting gRNA, to identify molecular...vitro" form. In this form, the CRISPR complex, consisting of a recombinant dCas9 protein and synthetic ...Yuno, and Hodaka Fujii. "An enChIP system for the analysis of bacterial genome functions." BMC research notes...expressing the 3x FLAG‐dC as9 protein for enCh IP analysis." Genes to cells 23.4 (2018): 318-325. PubMed ...

  6. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    Published
    Nov. 29, 2022, 2:15 p.m.
    ...primarily European reference genome could affect their analysis. Their work revealed that cell lines from individuals...information, Misek notes that in theory, if the DepMap analysis had an equal representation of every ancestry ...descent.Correcting the bias  While their thorough analysis uncovered some concerning biases within widely... team at Broad, they were able to change their analysis to use a corrected genome map. Now, any data that...

  7. Tips from the Repository Trenches: Using Barcodes to Track Samples

    Type
    Blog Post
    Published
    Oct. 6, 2015, 2:30 p.m.
    ...across oceans and continents. Each deposit may consist of anywhere from one to hundreds of samples, with...plasmids contain either an ampicillin or kanamycin resistance gene. Every step along the way involves good ...your lab ensure that sample ID’s are unique, consistently labeled, and that numbers are not switched around... with plasmid transformation, preparation, and analysis from our protocols and molecular biology reference...

  8. Plasmids 101: Protein tags

    Type
    Blog Post
    Published
    Dec. 11, 2014, 4:26 p.m.
    ...purification with glutathione, can also protect against proteolysis. That’s one form of instability. Prokaryotes ...purification with glutathione; protects against proteolysis, but may reduce solubililty HA YPYDVPDYA or...EINAPTDGKV EKVLVKERDA VQGGQGLIKI GVHHHHHH  9 Combo Consists of a bacterially derived in-vivo biotinylation...KEFLDANLAG SGSGHMHHHH HHSSGLVPRG  12 Solubililty Assists in proper folding V5 GKPIPNPLLGLDST   1.4 Affinity...

  9. Your Lentiviral Plasmid FAQs Answered

    Type
    Blog Post
    Published
    April 23, 2014, 1:08 p.m.
    ...1]; and 2) the potential for oncogenesis through insertional mutagenesis. These risks are largely based...differ in two ways. Firstly, 2nd generation systems consist of an envelope plasmid and a packaging plasmid ...the deletion reduces the risk of insertional mutagenesis by limiting the interaction between viral LTR...

  10. Viral Production at Addgene

    Type
    Blog Post
    Published
    Oct. 24, 2023, 1:15 p.m.
    ...preparations are subjected to polyacrylamide gel electrophoresis (PAGE) followed by silver staining or SYPRO...purified and non-purified AAV subjected to gel electrophoresis. Viral capsid proteins VP1, VP2, and VP3 are...preparation shows that the vast majority of the vectors consist of full particles (white arrowhead) relative to...transfection. PCR products are analyzed by gel electrophoresis to ensure the correct banding pattern and sizes...

  11. Getting the Most from Your Lentiviral Transduction

    Type
    Blog Post
    Published
    April 7, 2016, 2:30 p.m.
    ...other cell lines, or infected personnel and can persist undetected; unlike infections with larger microbes...al. found that lentivirus produced at pH 6 are resistant to multiple freeze thaw cycles. Consequently, ... to ensure the best possible transduction. Be consistent Once your lab establishes the best conditions...for your cell lines and virus properly and be consistent when following protocols. Need Virus? Check out...

  12. Important Considerations in Optogenetics Behavioral Experiments

    Type
    Blog Post
    Published
    Oct. 1, 2015, 2:30 p.m.
    ...rudimentary, but before you do any type of behavior analysis, simply take a few animals and stimulate them ...optogenetics experiment is contained within the hypothesis you are testing. In other words, your optogenetic...explained primarily in two different ways 1) your hypothesis is wrong or 2) there is some problem with the...optogenetic tools derived from direct comparative analysis of microbial opsins." Nature methods 9.2 (2012...

  13. Fluorescent Proteins 101: GFP Fusion Proteins - Making the Right Connection

    Type
    Blog Post
    Published
    April 9, 2019, 1:13 p.m.
    ...contributed by guest blogger Joachim Goedart, an assistant professor at the Section of Molecular Cytology...coded by the multiple cloning site and therefore consist of random amino acid sequences or (ii) designed... an inert, unstructured peptide and therefore consisted of glycines, serines and other small, non-aliphatic...blogger, Joachim Goedhart! Joachim Goedhart is an assistant professor at the Section of Molecular Cytology...

  14. Plasmids 101: Biotinylation

    Type
    Blog Post
    Published
    Nov. 15, 2018, 1:50 p.m.
    ... are essential for processes such as gluconeogenesis, lipogenesis, amino acid metabolism and energy transduction...Kresege et al., 2004). Avidin is a glycoprotein that consists of four identical subunits that can each bind ...non-selectively label DNA and RNA. Photobiotin acetate consists of a biotin molecule attached to a photoreactive...

  15. Hot Plasmids: Summer 2024

    Type
    Blog Post
    Published
    Aug. 27, 2024, 1:15 p.m.
    ...loss. Ci Ji Lim’s lab found that adding Late Embryogenesis Abundant (LEA) proteins, small proteins that... by helical LEAs. C-E) cryo-EM single-particle analysis of PRC2 with RvLEAMshort (1:6 molar ratio) with...s other recent deposit builds on Prime-editing-Assisted Site-Specific Integrase Gene Editing (PASSIGE)...CRISPRoff system — a programmable epigenetic editor consisting of dCas9 fused to a DNA methyltransferase — that...

  16. CRISPR 101: Validating Your Genome Edit

    Type
    Blog Post
    Published
    Nov. 3, 2022, 12:15 p.m.
    ...of WT and edited cell lines. Then decomposition analysis is performed yielding a graph of edits in the ... Fig. 3: Restriction enzyme screening for resistivity to digestion, indicating targeted edits. ...solely for restriction enzyme sensitivity or resistivity, and thus be a marker for edited cells. As with...Software such as CRISPResso can help with the data analysis. If NGS is not an option, you can still detect...

  17. Treating Muscular Dystrophy with CRISPR Gene Editing

    Type
    Blog Post
    Published
    Jan. 26, 2016, 3:30 p.m.
    ...off-target editing. In order for the therapy to persist over time, stem cell editing would be highly desirable...improved, with vastly reduced inflammation and necrosis compared to unedited animals. In grip strength...approach, increasing the possibility that edits will persist long-term. In accordance with this result, Nelson...CRISPR Could Be Used to Treat Cataracts and Cystic Fibrosis Read How Homology Directed Repair is Used to Edit...

  18. How to Design Your gRNA for CRISPR Genome Editing

    Type
    Blog Post
    Published
    Sept. 24, 2020, 1:15 p.m.
    ..., it would be unwise to make conclusions on the basis of the activity of a single gRNA, and thus diversity...on careful experimental design, execution, and analysis. Many thanks to our guest blogger John Doench....2017.97 Hanna RE, Doench JG (2020) Design and analysis of CRISPR–Cas experiments. Nat Biotechnol 38:813...https://doi.org/10.1038/s41576-018-0059-1 Veres A, Gosis BS, Ding Q, Collins R, Ragavendran A, Brand H, Erdin...

  19. Plasmids 101: Repressible Promoters

    Type
    Blog Post
    Published
    Dec. 20, 2022, 2:15 p.m.
    ...systems. Binary systems are genetic tools that consist of two parts, both of which must be present for...allows them to be used together for complex genetic analysis in Drosophila or for precise control over multiple...Desplan. “Power tools for gene expression and clonal analysis in Drosophila.” Nat Methods 9(1) (2011): 47-55...transgene expression, lineage tracing, and mosaic analysis.” Cell 141(3) (2010): 536-48. PubMed PMID: 20434990...

  20. Transferable Skills Guide: Teamwork

    Type
    Blog Post
    Published
    Nov. 28, 2017, 2 p.m.
    ...of a senior thesis), I knew early on that I wanted to work with a partner. WPI’s emphasis on teamwork ...decision about something? Fix a problem? Test a hypothesis? Whether you officially call it a “project team... risky shots at the goal rather than going for assists that are more likely to lead to scores. That player...

  21. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    Published
    March 21, 2017, 2:30 p.m.
    ...the Moloney Murine leukemia virus (MoMLV) and consisted of two plasmids: the packaging plasmid containing...generation. The 1st generation of lentiviral vectors consisted of three plasmids encoding 1) the lentiviral vector...Efficient gene transfer Potential for insertional mutagenesis: Replication-incompetent lentiviruses with human... from one batch to another. For experimental consistency it’s a good idea to purify and titer your virus...

  22. An “elegans” Approach to Better CRISPR/Cas9 Editing Efficiency

    Type
    Blog Post
    Published
    Dec. 17, 2020, 1:30 p.m.
    ...post was contributed by Jordan Ward who is an assistant professor at the University of California, Santa...elegans relied on selective markers such as drug resistance (Chen et al., 2013), fluorescence (GFP; Tzur ...Thank you to our guest blogger! Jordan Ward is an assistant professor at the University of California, Santa...on gene regulation in nematode molting and spermatogenesis. His group also builds tools to improve genome...

  23. CRISPR 101: Cytosine Transversion Editors

    Type
    Blog Post
    Published
    March 25, 2025, 1:15 p.m.
    .... Like other ABEs, the editor they were using consisted of Cas9 fused to an engineered adenosine deaminase...tool for predicting CGBE editing outcomes and assisting sgRNA selection is available at http://www.sunlab.fun...A., Yang, D., Mok, B., Replogle, J. M., Xu, A., Sisley, T. A., Weissman, J. S., Adamson, B., & Liu, D....developed using CRISPRi screens, target-library analysis, and machine learning. Nature Biotechnology, 39...

  24. Advice on Career Paths and the Green Card Process for International Researchers and Entrepreneurs

    Type
    Blog Post
    Published
    Dec. 20, 2017, 2:33 p.m.
    ...a prestigious institution: This criterion is consistently under scrutiny. USCIS wants you to prove that... is very important in this case.  You can still assist your PI in the peer review process, but remember...love of science and technology, Rachel is proud to assist companies and foreign nationals working in the ...dedicated to the practice of US immigration law and assists individuals, researchers, entrepreneurs, companies...

  25. Editor's Choice, August 2016

    Type
    Blog Post
    Published
    Sept. 2, 2016, 1:32 p.m.
    ...optimize and troubleshoot the system. Site Directed Mutagenesis by PCR & Single Base Editing with CRISPR For ...technology may make some uses of Site Directed Mutagenesis by PCR obsolete! Honorable mentions Of course... in chronological order here: Site Directed Mutagenesis by PCR Google Forums Round Up: First Impressions...

  26. Who Gives a Tweet? 9 Facts About Scientists on Twitter

    Type
    Blog Post
    Published
    June 26, 2017, 1:31 p.m.
    ...this article: “A systematic identification and analysis of scientists on Twitter.” This paper addresses..., or are friends, are included in this cluster analysis. The closer two dots are to each other, the more... C. R. (2017). A systematic identification and analysis of scientists on Twitter. Plos One,12(4). doi:...

  27. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    Published
    Feb. 13, 2025, 2:15 p.m.
    ... than point modifications. CRISPR base editing persists through multiple cell divisions, indicating that...specificity. In this first, the Liu lab used phage-assisted evolution selection systems to generate ABE8e(... a similar approach. Dali Li’s lab’s A&C-BEmax consists of a fusion of both cytidine and adenosine deaminases... E., Doudna, J. A., & Liu, D. R. (2020). Phage-assisted evolution of an adenine base editor with improved...

  28. Sensing Neuronal Dopamine

    Type
    Blog Post
    Published
    July 30, 2019, 12:59 p.m.
    ...dopamine. Methods for measuring Dopamine In vivo microdialysis The gold standard method of measuring DA and...neurotransmitters in the brain has been in vivo microdialysis of extracellular fluid (Darvesh et al., 2011...References Darvesh, Altaf S., et al. "In vivo brain microdialysis: advances in neuropsychopharmacology and drug...

  29. FastCloning: A Newer, Simpler Cloning Technique

    Type
    Blog Post
    Published
    Nov. 1, 2016, 2:30 p.m.
    ... these techniques and is a multi-step process consisting of digestion, purification, ligation, and transformation... 1. Celie, Patrick HN, Annabel HA Parret, and Anastassis Perrakis. "Recombinant cloning strategies for...Plasmid Cloning by PCR Perform Site Directed Mutagenesis by PCR Catch up on Your Plasmid Background with...

  30. Sleeping Beauty Awakens for Genome Engineering

    Type
    Blog Post
    Published
    June 30, 2015, 2 p.m.
    ...social and regulatory reasons. 1.) The transposon consists of a mirrored set of inverted repeats (green) ...1) an SB transposase, the enzyme required for catalysis of transposition; and 2) a transposon containing...determined by transposition assays of antibiotic-resistance genes conducted in HeLa cells). In their Nature...

  31. CRISPR Challenges: Standardization and Homology Directed Repair

    Type
    Blog Post
    Published
    Feb. 14, 2018, 2:36 p.m.
    ...much more about cellular DNA repair pathways. An emphasis on cellular repair post-CRISPR cleavage When I...Ribonucleoprotein (RNP) Delivery The RNP delivery method consistently leads to high on-target editing and low off-...strategies activate different HDR sub-pathways like synthesis-dependent strand annealing (SDSA) and single-strand...

  32. In Vivo Biotinylation of Bacterial Fusion Proteins

    Type
    Blog Post
    Published
    Jan. 25, 2018, 2:09 p.m.
    ...GST-glutathione. Biotin-based immobilization for kinetic analysis Proteolytic removal of the fusion protein could...could provide a more relevant kinetic analysis, but the trick is to utilize a second immobilization method... purification strategy outlined here provides a basis to determine binding kinetics using a multitude ...

  33. Single-cell tracking of lineage and identity with CellTag

    Type
    Blog Post
    Published
    May 9, 2019, 1:15 p.m.
    ...labels cells with heritable barcodes. Barcodes consist of an 8bp variable region and a 6 bp fixed region... a sample of cells was collected for scRNA-seq analysis and the remaining cells were maintained in culture...detailed CellTag protocol on protocols.io, code and analysis tutorials on GitHub, and a CellTag lineage data...

  34. Bright Monomeric Fluorescent Proteins: mNeonGreen, mTFP1, and mWasabi

    Type
    Blog Post
    Published
    April 25, 2019, 3:01 p.m.
    ...was then screened through successive rounds of mutagenesis for the brightest teal fluorescent proteins that...scientists from Florida State University, they used a mutagenesis approach and found that specific amino acid changes...protein localized to their expected locations. An analysis of mWasabi gave similar results. We hope this ...

  35. Three Tips for Preventing Viral Plasmid Recombination in Your Samples

    Type
    Blog Post
    Published
    Sept. 14, 2021, 1:15 p.m.
    ...culture, only the plasmid with the antibiotic resistance gene and origin will replicate, and the transgene...recombined vector backbone contains the antibiotic resistance gene (AmpR) and origins of replication (ori and...of bacterial plasmid DNA: electron microscopic analysis of in vitro intramolecular recombination. Proceedings...

  36. Science Careers: Unruly Interests Feed Many Paths

    Type
    Blog Post
    Published
    Sept. 26, 2017, 12:12 p.m.
    ...replication and gene transcription during vertebrate embryogenesis. My interests, though, have proven unruly. Instead... at Science magazine. That one I just could not resist applying for. Not that I knew anything about being...like word choice, punctuation, and stylistic consistency. If you’ve ever doubted the importance of copy...

  37. Plasmids 101: TOPO Cloning

    Type
    Blog Post
    Published
    Oct. 27, 2016, 2:30 p.m.
    ... Blog Perform Site Directed Mutagenesis by PCR Use REPLACR Mutatgenesis to Easily Generate Insertion ...approach to molecular cloning and polynucleotide synthesis using vaccinia DNA topoisomerase. Shuman S. J ...

  38. Plasmid Cloning by PCR

    Type
    Blog Post
    Published
    March 29, 2016, 2:30 p.m.
    ...molecular cloning consist of: Leader Sequence: Extra base pairs on the 5' end of the primer assist with restriction...selecting restriction sites, you should use a DNA analysis tool, such as Addgene’s Sequence Analyzer, to ...

  39. Creating accessible biology activities in schools with BioBits

    Type
    Blog Post
    Published
    Aug. 2, 2018, 12:56 p.m.
    ... way into high school classrooms, like gel electrophoresis to separate DNA or transforming cells with ...practice with practical lab skills like developing a hypothesis, precisions pipetting, and good record keeping...in functional genomics research Learn how the Arabidopsis research center brings plant science to life ...

  40. "Build Your Plasmid": The Game - Play to Teach

    Type
    Blog Post
    Published
    July 17, 2018, 12:23 p.m.
    ...about Synthetic Biology Find Arabidopsis educational resources at the Arabidopsis Biological Resource Center...Promoters, Inserts, Tags, Origins of Replication, Resistance gene and Selection Markers) and one “Transfer...
Showing: 161 - 200 of 608 results