We narrowed to 983 results for: ran
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TypeBlog Post...fragments of these transcripts. I remember when small RNA was just something we ran off the bottom of ...’t give tRNAs much thought. High throughput transcriptome sequencing methods including work done by the...hypothesized that life started with RNA and a simple cell membrane. His recent work shows that they are making fairly...knew…I had barely heard of these). These single stranded, covalently closed RNAs seem to be conserved in...
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Story of a SynBio Startup: RevBio's Epiphany (or Lack Thereof)
TypeBlog Post...that produce GMO food plants. When a press piece ran on the detector plants, we expected to see that same...below. We’re happy to answer questions about this transition (so far) and what we’ve learned. Thank you ... -
Design Tips for Prime Editing
TypeBlog Post...doi.org/10.1038/s41587-021-01039-7 Nishimasu, H., Ran, F. A., Hsu, P. D., Konermann, S., Shehata, S. I....nicking of the unedited strand and encourage the cell to use the edited strand as a template during DNA...on a fusion enzyme of Cas9 nickase + reverse transcriptase and a prime editing guide RNA (pegRNA). The ...) complementary to the genome and a reverse transcriptase (RT) template including the desired edit. ...although sequences with G/C content outside this range can still be optimized (Anzalone et al., 2019). .... Test different lengths of reverse transcriptase template, starting with about 10–16 nucleotides. For...pegRNA primer binding site (PBS) and reverse transcriptase template (RTT). Created with BioRender.com. ... -
Bioinformatics at Addgene
TypeBlog Post... same output regardless of who or which computer ran the process. Another design choice we appreciated...automating the bioinformatics steps needed to transform the raw sequencing data into a complete plasmid...appropriate place in our workflow without having to translate or convert programs to a uniform language. The...concepts and vocabulary of Nextflow so that we could translate our pipeline steps into processes and then connect... -
DNA Purification Without a Kit
TypeBlog Post...just make their own buffers when the kits buffers ran out. Homemade buffers performed the same as those...rather a glass powder slurry that has a milk-like appearance. To make DNA stick to the glass particles in ...the Patterson Protocol was shown to purify DNA ranging in size from 100 bp to 48 kb with high yields and...could interfere with interpreting the results of transfection assays. Go forth and perform your DNA extractions... -
Immunology Research Plasmids and Resources
TypeCollection...- CHGA chromogranin A (parathyroid secretory protein 1) CGA CHGB chromogranin B (secretogranin 1) SCG1...superfamily, member 11 CD254, ODF, OPGL, OPTB2, RANKL, TRANCE, hRANKL2, sOdf TNFSF12 tumor necrosis factor (ligand... perforin and granzyme B, two enzymes that create pores within an infected cell membrane, triggering cell...nuclear transcription factor Y, alpha CBF-A, CBF-B, FLJ11236, HAP2, NF-YA NFYB nuclear transcription factor...RAET1G retinoic acid early transcript 1G ULBP5 RAET1L retinoic acid early transcript 1L - RELB v-rel reticuloendotheliosis...(sodium/bile acid cotransporter family), member 2 ASBT, ISBT, NTCP2 TAP1 transporter 1, ATP-binding cassette... -
Tetracycline Inducible Expression
TypeCollection...tetracycline-controlled transactivator (fusion of TetR with VP16 transcriptional activation domain); rtTA...tetracycline-controlled transactivator (fusion of mutated TetR with VP16 transcriptional activation domain)... integrate a transactivator under a tissue-specific promoter and administer the transgene separately. ...easier to implement and ensures that all transfected or transduced cells express both components in the same...generations of transactivators and promoters are generally cross-compatible. Your choice of transactivator, promoter...promoter, and type of transduction (transient or stable) may depend on the desired sensitivity, level of...Return to Top Transactivators (tTA or rtTA) Find a construct that expresses the transactivator for your tetracycline... -
Fluorescent Protein Guide: Subcellular Localization
TypeCollection...37351 pQC membrane TdTomato IX Membranes Palmitoylation tdTomato Connie Cepko 22479 FUmGW Membranes Palmitoylation...-1 mGold2t Francois St-Pierre 231774 pCMV-mGold2s-CAF1-C-10 Nucleus CAF-1 mGold2s Francois St-Pierre 189775...pCAG-mGFP Membranes Palmitoylation sequence from GAP43 EGFP Connie Cepko 61099 Lck-GFP Membranes Palmitoylation...outer membrane TOMM20 mCherry* Michael Davidson 118736 pBOB-CARMIL2 BH domain-GFP Plasma membrane CARMIL2...Plasma membrane CARMIL2 BH domain tdTomato John Cooper 189771 pLCK-mScarlet3_C1 Plasma membrane Lck N-...mGold2t Francois St-Pierre 231772 pCMV-mGold2s-Actin-C-18 Actin filaments beta-Actin mGold2s Francois St-Pierre...EGFP Carol Otey 31923 pPaxillin-PSmOrange Focal adhesions Paxillin PSmOrange Vladislav Verkhusha 36991 Desmoglein2... -
Fluorescent Protein Guide: Biosensors
TypeCollection...Wolf Frommer Calcium Orange fluorescent calcium ion indicator OCaMP A sensitive orange fluorescent calcium...for glutamatergic neurotransmission Targeted sensors for glutamatergic neurotransmission. eLife. 2023 Jan...trafficking to the plasma membrane Acute disruption of the synaptic vesicle membrane protein synaptotagmin...measurements of membrane charges A FRET sensor enables quantitative measurements of membrane charges in live...847-56. David Raible pH Red or Orange pH biosensors (pHuji and pHoran4) pHuji, a pH-sensitive red fluorescent...Proteins: Biosensors Metal Ions Small Molecules Neurotransmitters Cell Physiology Browse All Other Reporters...protein (cpFP) or fluorescence resonance energy transfer (FRET) efficiency of a pair of fluorescent proteins... -
Plan Your Experiment
TypeCollection...fused to transcriptional activators that need to be within a given range of the transcription start site... interfere with transcription elongation. CRISPR Activation : Target the transcription start site. CRISPRa... perform by chemical transfection or electroporation. Expression can be transient, or you can generate...efficiently transfect. However, note that this method is most useful for cell lines that you can transfect at ...into cells. These methods are more transient than plasmid transfection or viral vectors, reducing the risk...your target cells are not transcribing DNA to mRNA, nor do they need to translate mRNA to protein. Instead...is helpful in cells that are difficult to transfect, transduce, or may not express common promoters. Browse... -
Open Enzyme Collection
TypeCollection...pOpen-EcoRIM M.EcoRI DNA methyltransferase 165517 pOpen-Eco31IA Eco31IA DNA methyltransferase 165518 pOpen-NotIM...pOpen-NotIM M.NotI DNA methyltransferase 165527 pOpen-Eco31IB Eco31IB DNA methyltransferase 165536 pOpen-PstIM...pOpen-PstIM M.PstI DNA methyltransferase 165538 pOpen-SfiIM M.SfiI 165538 DNA methyltransferase 165547 pOpen-SapIM1...pOpen-SapIM1 M1.SapI DNA methyltransferase 165557 pOpen-SapIM2 M2.SapI DNA methyltransferase 165558 pOpen-EcoRVM...pOpen-EcoRVM M.EcoRV DNA methyltransferase 165577 pOpen-XbaIM M.XbaI DNA methyltransferase 165578 pOpen-NcoIM ...32 single-stranded DNA-binding protein 165549 pOpen-ETSSB Extreme Thermostable Single-Stranded DNA-Binding...Polymerases Restriction Endonucleases Reverse Transcriptases RNA Polymerases Other Enzymes Additional Resources... -
Luciferase Plasmid Collection
TypeCollection...Bioluminescence Resonance Energy Transfer (BRET) Similar to FRET, energy transfer from the luciferase reaction...can be used to study processes beyond transcription and translation. Splicing reporters from Younis et al...bioluminescence found in a variety of organisms, ranging from bacteria to aquatic animals to insects. Luciferase...photons. Its robust signal, sensitivity, wide dynamic range, and resistance to photobleaching have made luciferase...Regulatory elements such as promoters, enhancers and untranslated regions, or shRNA or CRISPR target sites can... protein half-lives, making them useful as transcriptional reporters, but are less-than-ideal protein ...Vector for investigating regions controlling transcription Debrya Groskreutz 18760 MSCV IRES Luciferase... -
Fluorescent Protein Guide: Empty Backbones
TypeCollection... Monomer pET mOrange LIC cloning vector (u-mOrange) - Bacterial Expression pCS2+8CmOrange - Zebrafish/...urchin pCS2+8NmOrange - Zebrafish/Xenopus/Worm/Sea urchin mOrange-N1 - Mammalian Expression mOrange-C1 - Mammalian...Expression mOrange-pBAD - Bacterial Expression mOrange2 549 565 35 6.5 4.5 h Monomer mOrange2-N1 - Mammalian...Expression LSSmOrange 437 572 23 5.7 2.3 h Monomer pLSSmOrange-C1 - Mammalian Expression pLSSmOrange-N1 - Mammalian...Structure Plasmids mKusabira-Orange (mKO) 548 559 31 5 4.5 h Monomer pQC mKorange IX - Mammalian Expression...Expression mKusabira Orange-C1 (mKO) - Mammalian Expression mKO-pBAD - Bacterial Expression mOrange 548 562 49 ...Mammalian Expression mOrange2-C1 - Mammalian Expression mOrange2-pBAD - Bacterial Expression mKO2 551 565... -
Lentivirus Plasmids
TypeCollection...divided among multiple plasmids: Transfer plasmid — contains transgene, sgRNA, or shRNA of interest flanked...envelope, and transfer plasmids Viral... Viral Plasmids Lentiviral Plasmids Lentiviral Plasmids Transfer Plasmids Packaging...types of viruses used in research. Lentivirus can transduce both dividing and non-dividing cells without a...stably into the host genome, enabling long-term transgene expression. Wild-type lentiviruses have been engineered...third-generation lentiviral packaging, envelope, and transfer plasmids available at Addgene. Read our Lentiviral...safety. Lentiviral Vector Guide Popular Lentiviral Transfer Plasmids Looking for lentiviral plasmids to express... -
University of Florida Serotype Testing Panel for the Eye and Brain
TypeCollection...enhanced transduction of the retina following subretinal administration and enhanced transduction and spread...cite Boye, et al. 2016. Impact of Heparan Sulfate Binding on Transduction of Retina by Recombinant Adeno-... from the University of Florida for enhanced transduction of the eye and other tissues. As part of our...Y444F) serotype is demonstrated to have enhanced transduction of the retina following intravitreal injection... AAV2(Y444F) serotype also displays enhanced transduction of the trabecular meshwork following intracameral...trpYF) The AAV2(trpYF) serotype shows enhanced transduction of the retina following intravitreal injection...macaque. This serotype also demonstrates enhanced transduction of cells in culture for ex-vivo conditioning... -
Plasmids for Stem Cell Research
TypeCollection...reprogramming factors Efficient germ-line transmission obtained with transgene-free induced pluripotent stem cells...Reprogramming Plasmids Differentiation and Transdifferentiation Plasmids Stem Cell Research Plasmids Stem...differentiated somatic cell into another via transdifferentiation. This method of cell lineage conversion ...technology. Browse Addgene's collection of Transdifferentiation Plasmids below. Many of the techniques for... for reprogramming, differentiation, and transdifferentiation utilize plasmids to increase expression ...human Klf4, Oct4, c-Myc, and Sox2 as VP-16 transcriptional activating fusions in single lentiviral vectors...human iPSC generation OCT4 and SOX2 Work as Transcriptional Activators in Reprogramming Human Fibroblasts... -
Viral Vector Guides and Plasmids
TypeCollection...packaging capacity, high transduction efficiency, and high but transient transgene expression.They can induce...130 nm Dividing Yes Moderate AAV Transient or Stable* Single-stranded linear DNA ∼4.5 kb 18–26 nm Dividing.../Non-dividing No* Very Low Adenoviral Transient Double-stranded linear DNA 5–36 kb 105 nm Dividing/Non-dividing...of viral vectors and find viral production and transfer plasmids for a wide array of research purposes...are versatile tools because of their ability to transduce dividing and non-dividing cells. These virusal...immunogencity. These viral vectors can direct long-term transgene expression, but generally don't integrate into...just retroviral vectors, have a relatively high transduction efficiency and only infect dividing cells. They... -
Empty Backbones - Choosing Your Perfect Plasmid Backbone
TypeCollection...expression Find yeast transformation plasmids with G418 selection Find nematode transgenesis plasmids with Neomycin...Backbones Mammalian CMV, SV40, EF1a, CAG, Ubc Transient expression - Gradia Lab plasmids for mammalian...gene with N-terminal GST tag and TEV cleavage Transient expression - Gradia Lab plasmids for bacterial...This means that the final product should be translated as a single string of amino acids that preserves...with N-terminal Myc tag and attB for Drosophila transgenesis His Epitope tag pEZYmyc-His - C-terminal ...3Myc-EYFP-HA-His6 and IRES-dependent ECFP-HA-His6 translation for mammalian expression pDest-527 - N-terminal...EBFP2 blue fluorescent protein 35S-eGFP-nosT - Transient expression vector for eGFP in plants Find FP tagging... -
Fluorescent Proteins: FRET
TypeCollection...pBAD , pLSSmOrange-mKate2 LSSmOrange* mScarlet3 437 0.45 592 104,000 0.75 5.8 4.3 pLSSmOrange-N1 , pmScarlet3...Resonance Energy Transfer (FRET) is a process by which energy is non-radiatively transferred from an excited...2.9 mVenus N1 , mKO2-N1 mOrange2 mCherry 549 0.60 610 72,000 0.22 6.1 4.4 mOrange2-N1 , mCherry2-N1 miRFP670...-N1 , pmiRFP720-N1 LSSmOrange* mKate2 437 0.45 633 62,500 0.40 5.6 3.7 pLSSmOrange-N1 , mKate1.31-pBAD...energy transfer mCerulean, mVenus, and Amber (mVenus Y67C) constructs for measuring energy transfer to multiple...FLIM) or FLIM-FRET, where the increased energy transfer through FRET reduces the donor lifetime. FRET ...2006) Quantitative understanding of the energy transfer between fluorescent proteins connected via flexible... -
Retrovirus Plasmids
TypeCollection...into multiple plasmids for safety: Transfer plasmid — contains transgene, sgRNA, or shRNA of interest flanked... the transgene fused to eGFP. Hans Clevers 18760 MSCV IRES Luciferase MSCV Plasmid for transgene expression...and transfer plasmids. Viral... Viral Plasmids Gamma-Retroviral Plasmids Gamma-Retroviral Plasmids Transfer Plasmids...encoded on RNA. The genome includes a reverse transcriptase and an integrase to insert the viral genome ...can take advantage of this to stably integrate transgenes of interest into a variety of cell types. Wild-type...collection highlights popular gamma-retroviral transfer, packaging, and envelope plasmids. Read our Gamma-retroviral...