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Showing: 21 - 40 of 49 results

  1. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...by Limiting Dilution You may also like... Viral Vector Guides Virus Blog Posts Addgene Protocols Viral...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette CO 2 incubator Pipet controller...96-well plate, seed one of the corner wells with ~1000 cells. This will help you focus the microscope later... incubating the cells. Pro-Tip Use the well with 1000 cells to focus the microscope, then you can easily... plasmid #52962 ) and is described in Improved vectors and genome-wide libraries for CRISPR screening....

  2. Colony Formation Titering Assay

    Type
    Protocol
    ...Assay for Lentivirus You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Viral...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Ice bucket CO 2 incubator...the indicated serial dilutions of the lentiviral vector pRosetta . The following day, cells were treated...the indicated serial dilutions of the lentiviral vector pHAGE EF1α dCas9-KRAB . The following day, cells...

  3. Fluorescent Protein Guide: Empty Backbones

    Type
    Collection
    ...Expression (this is a bicistronic vector rather than a fusion protein vector) ZsGreen1-N1 - Mammalian Expression...exctinction coefficient and quantum yield divided by 1000. pKa is the pH at which fluorescence intensity is...93 5.7 10 min Monomer pCS2+mNeonGreen-C Cloning Vector - Mammalian Expression Jump to Top Yellow Protein...5.7 Monomer (A206K) pcDNA3 mCitrine LIC cloning vector (6C) - Mammalian Expression pCS2+8CmCitrine - Zebrafish...Xenopus/C.elegans/Sea urchin pET mCitrine LIC cloning vector (u-mCitrine) - Bacterial Expression mCitrine-N1...562 49 6.5 2.5 hr Monomer pET mOrange LIC cloning vector (u-mOrange) - Bacterial Expression pCS2+8CmOrange... - Mammalian Expression pCS2+mScarlet-C Cloning Vector - Mammalian Expression mScarlet-I 569 593 57 5.4...

  4. Using CRISPR/Cas9 to Edit Disease Out of the Genome

    Type
    Blog Post
    ...control the CRISPR system activity.  Using an AAV vector may help with size limitations. In addition, non-viral...& Parkinsonism. https://doi.org/10.4172/2161-0460.1000439 6. Wu et al. 2020. . “Advances in CRISPR/Cas-based...

  5. General Transfection

    Type
    Protocol
    ...General Transfection You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Viral...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Ice bucket CO 2 incubator...solution through a 0.22 µm membrane. Aliquot 500–1000 µL into sterile tubes. Store the tubes at -80 °C...

  6. Lentivirus ddPCR Titration

    Type
    Protocol
    ...ddPCR) for lentivirus vector titration.... Protocols ddPCR Titration of Lentivirus Vectors ddPCR Titration of...of Lentivirus Vectors You may also like... Lentivirus Colony Formation Titration Assay Lentivirus Fluorescence...droplet digital PCR (ddPCR) to titer lentivirus vectors. This protocol specifically uses primers and probes...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette 2–50 µL multichannel pipette...

  7. Tips for Using FRET in Your Experiments

    Type
    Blog Post
    ...pGWF1 Cyan & Yellow Bacterial Gateway-compatible vector to express a gene of interest fused between ECFP...Bioessays. 2012 May; 34(5):341-50. doi: 10.1002/bies.201100098. Sekar RB, Periasamy A. Fluorescence resonance...

  8. AAV Production in HEK293 Cells

    Type
    Protocol
    ...Cells You may also like... AAV Viral Service Viral Vector Guides Virus Blog Posts Molecular Biology Reference...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette Hemocytometer or cell ...system, Nalgene, 565-0010 (or for larger volumes 1000 mL 0.45 µm Rapid-Flow PES Filtration Unit, Nalgene...solution through a 0.22 μm membrane. Aliquot 500–1000 μL into sterile tubes. Store the tubes at -80 °C... deionized water and adjust to a final volume of 1000 mL. Stir at RT until fully dissolved. Pro-Tip This...

  9. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    ...more permanent knockdown, consider using shRNA vectors instead. With both siRNA and shRNA, it can be difficult...and flow cytometry. F1000Research, 13, 817. https://doi.org/10.12688/f1000research.154034.1 Laflamme, ...synaptotagmin 1 page. The data produced is available on the F1000 gateway and Zenodo community.  Knockdown approach...

  10. Protocol - Bacterial Transformation

    Type
    Protocol
    ...approximately 10-fold lower for ligation of inserts to vectors than for an intact control plasmid. Incubate the...when using highly competent cells. If you used 100-1000 ng of total DNA in a ligation you will often get...

  11. AAV Titration by qPCR Using SYBR Green Technology

    Type
    Protocol
    ...SYBR Green Technology You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Introduction...the titer of samples obtained from academic viral vector cores. Our titers were similar to those reported...channel pipette 20–200 µL single channel pipette 200–1000 µL single channel pipette 1–10 µL multichannel pipette...95 uL 20X 200X Dilution 3 20 uL Dil. 2 80 uL 5X 1000X Dilution 4 20 uL Dil. 3 80 uL 5X 5000X Dilution ...

  13. Promoters

    Type
    Guide
    ... Promoters are a vital component of expression vectors because they control the binding of RNA polymerase...interest is expressed. Summary Promoters are about 100-1000 base pairs long and are adjacent and typically upstream...

  14. Plasmids 101: Dimers and Multimers

    Type
    Blog Post
    ... JA, Carnes AE, Hodgson CP. Plasmid DNA vaccine vector design: impact on efficacy, safety and upstream...Electrophoresis. 2010 Jul;31(14):2436-41. doi: 10.1002/elps.201000061. PMID: 20589859. Levene SD. Analysis of DNA topoisomers...

  16. Penn Vector Core Partnership with Addgene

    Type
    Collection
    ...Viral Vector Packaging Service Penn Vector Core Transfer Viral Vector Packaging Service: Penn Vector Core...University of Pennsylvania Vector Core (Link opens in a new window) (Penn Vector Core) and Addgene have partnered...provide high-quality AAV vectors to the academic research community. The Penn Vector Core will produce viral-based...viral-based vectors for distribution via Addgene. For a full list of Addgene's viral vectors See Addgene's...inventory , which includes all viral vectors transferred from the Penn Vector Core. Chemogenetics (including...a viral vector with the Penn catalog ID: Based on low demand, production of some viral vectors have been...See the list of discontinued viral vectors . The following vectors are now available for distribution ...

  17. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ... Ultracentrifugation You may also like... Viral Vector Guides Virus Blog Posts Mol Bio Protocols Introduction...and characterization of adeno-associated viral vectors," (Link opens in a new window) copyright (2006)...bring up the volume with formulation buffer. Use a P1000 to the bottom of the filter and pipette up/down ...

  18. Molecular Biology Reference

    Type
    Guide
    ...Commonly used cloning vectors include Gateway entry vectors and TOPO cloning vectors. If you are looking...combination of a DNA fragment with a plasmid or vector DNA backbone generates a recombinant DNA molecule...specifically engineered plasmids, commonly referred to as vectors, which have become possibly the most ubiquitous...facilitate the cloning of DNA fragments. Cloning vectors tend to be very simple, often containing only a...expression (for the purposes of gene study). Expression vectors must contain a promoter sequence, a transcription...the transcription process to stop. An expression vector can also include an enhancer sequence which increases... amount of protein or RNA produced. Expression vectors can drive expression in various cell types (mammalian...

  19. Optogenetics AAV Preps

    Type
    Collection
    ... Viral Vector Packaging Service AAV Optogenetics Viral Vector Packaging Service: Optogenetics... eligible AAV plasmids and we’ll make the viral vector prep you need. Or fill out our form to let us know...format. Please note this does not guarantee viral vector packaging service, but lets us know what viruses...GFP Syn ChR2/H134R GFP Constitutive 8, rg* Boyden 100054 pAAV.CAG.hChR2(H134R)-mCherry.WPRE.SV40 CAG ChR2...] CAG ChrimsonR tdTomato Flp dependent 8 Boyden 100049 pAAV.hSynap.ChETA(E123T/H134R)-eYFP.WPRE.hGH Syn...Syn ChETA EYFP Constitutive 1, 9 Deisseroth 100050 pAAV.CaMKIIa.ChETA(E123T/H134R)-eYFP.WPRE.hGH CaMKII... EYFP Constitutive 1, 5, 9, PHPeB Dimidschstein 100061 pAAV-Ef1a-DIO C1V1 (t/t)-TS-mCherry EF1a C1V1 (...

  20. CRISPR Pooled gRNA Libraries

    Type
    Collection
    ...-FokI Screen Pooled Libraries gRNAs Empty gRNA Vectors gRNAs By Species Mammalian Bacteria Drosophila ...TP53 Exon Mutation Libraries 229137–229140 Donor Vector Human Stiewe N/A Varies 1,910–3,034 per exon Human...Blast 1000000173 Knockout Mouse Liu 3rd 10 185,836 Mouse GeCKO v2 1000000052 (1 plasmid) 1000000053 (2 plasmid...Varies Bassik Mouse CRISPR Knockout Library 1000000121 — 1000000130 Knockout Mouse Bassik 3rd 10 Varies Berger...83978 1000000091 Activation Human Weissman 3rd 5 10 104,540 209,080 CRISPRa-v2 83996 1000000093 Activation...83969 1000000090 Inhibition Human Weissman 3rd 5 10 104,535 209,070 CRISPRi-v2 83987 1000000092 Inhibition...Birsoy 3rd 8 20,051 Human GeCKO v2 1000000048 (1 plasmid) 1000000049 (2 plasmid) Knockout Human Zhang 3rd...
Showing: 21 - 40 of 49 results