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  2. Plasmids 101: In-house NGS Plasmid Quality Control Now at Addgene

    Type
    Blog Post
    ...and last updated Apr 6, 2021 by Will Arnold. Introducing NGS in 2017 for all incoming plasmids was a big...Addgene - we had used Sanger sequencing for quality control for over ten years. Now, thanks to a generous donation...quantification of various intermediate steps, a sizing electrophoretic gel, and qPCR quantification of the final libraries...The first step we take when performing quality control of newly deposited plasmids is to align the NGS... by the depositing lab, we call these “quality control (QC) issues.” We then ask the depositing laboratory...

  3. Plasmids 101: Walkthrough of Addgene’s Snapgene-Powered Quality Control Process

    Type
    Blog Post
    ...talked a lot about the quality control process at Addgene by introducing our new sequencing partner seqWell...generation sequencing results to perform quality control on deposited plasmids. We’ve also talked about ...interface. We regularly use Snapgene for our quality control process because of its expansive feature library... main ways we use SnapGene for plasmid quality control are: To identify important plasmid features ...Identifying plasmid features While our quality control process for incoming plasmids can vary based on... quality control... plasmid quality control...

  4. Export Control Regulations Lifted for Plasmids Containing Rabies and VSV-G Genes

    Type
    Blog Post
    ...pUMVC (MuLV retroviral). Bob Weinberg pMD2.G VSV-G envelope expressing plasmid Didier Trono pVSV-G...plasmids. This is because in 2024 certain export control exceptions were made by the US Bureau of Industry...Industry and Security, pursuant to the international controls prescribed by the Australia Group, for these plasmids...Envelope protein for producing lentiviral and MuLV retroviral particles. Use in conjunction with a packaging...researchers in countries with applicable export control exceptions for rabies and VSV-G genes, plasmids...strive to keep our information updated, export controls and regulations are complex and may change suddenly...

  5. Teaching an Old DOG New Tricks: Controlling Protein Activity with GFP

    Type
    Blog Post
    ... just label cells. If GFP could be co-opted to control gene expression, he could then selectively manipulate...promoters. Moving to an in vivo system, Tang et al. electroporated T-DDOGs, GFP, and a reporter construct into...TdTomato, whose expression was absent without electroporated GFP. They then tested T-DDOGs with two mouse...robust and specific. When tested in retinal electroporation studies, DsRed reporter expression was induced...

  7. A New Optogenetic Tool Based on AraC Controls Gene Expression with Blue Light

    Type
    Blog Post
    ...Their system is based on the AraC protein that controls transcription from the PBAD promoter. “AraC is... promoter. While none of the constructs gave as strong a signal as with the wild-type AraC, it still reached... of interest, under the pBAD promoter, can be controlled with light without the need of any cloning. ...

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

    Type
    Blog Post
    ...an appropriate control? Many CRISPR screens use non-targeting guides as negative controls. Non-targeting...are well characterized and strategies exist to control for these faults. However, it’s still unclear if...screens and how best to design these screens to controls for flaws. Recently the Bassik Lab at Stanford...site in the genome. This means that while they control for many of the effects of gRNA and Cas9 expression...detecting Cas9 cutting. Safe targeting guides better control for non-specific toxicity than non-targeting guides... a coding region. Targeting these sites should control for the effects of guide expression and dsDNA breaks... the genome, and are commonly used as negative controls for genome-wide CRISPR screens; and 3) safe-targeting...

  10. Quick Guide to Working with Drosophila Part 2: Controlling Gene Expression in Flies with Gal4/UAS

    Type
    Blog Post
    ...systematically study gene function with temporal control and cell-type specificity! The Gal4/UAS system...you mate your fly line containing YFG under the control of the UAS with a fly line expressing Gal4 in your...promoter each time a new hypothesis is tested. Controlling multiple genes with Gal4/UAS system One can also...to drive expression of multiple genes under the control of UAS enhancer sequences. For example, if you ...them individually or simultaneously. Do note for control purposes that there are dosage effects. For example...

  11. Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens

    Type
    Blog Post
    ...for gene control over a 1,000-fold range. Weissman explained that there are two levels of control in the...technology has changed how scientists edit and control genes, but according to the Broad Institute's Silvana...screening In October, Jonathan Weissman’s lab introduced additional pooled CRISPR libraries, also available...abilities to turn genes on and off,” he said. “Some are strong and some more moderate; there is an allelic series...size of the backbone, transformations require electroporation. References & Further Reading: For more ...Epub 2014 Dec 10. Genome-Scale CRISPR-Mediated Control of Gene Repression and Activation. Gilbert LA, ...

  12. The Pleiades Promoter Project

    Type
    Collection
    ...CARTPT pEMS1492 intron-lacZ/NLS Ple22 CCKBR pEMS1493 intron-lacZ/NLS Ple23 CCKBR pEMS1494 intron-lacZ/NLS Ple24...CCKBR pEMS1495 intron-lacZ/NLS Ple25 CCKBR pEMS1496 intron-lacZ/NLS Ple26 CCL27 pEMS1497 intron-lacZ/NLS Ple27...CCL27 pEMS1498 intron-lacZ/NLS Ple28 CCL27 pEMS1499 intron-lacZ/NLS Ple29 CCL27 pEMS1500 intron-lacZ/NLS Ple30...CLDN5 pEMS1504 intron-lacZ/NLS Ple34 CLDN5 pEMS1505 intron-lacZ/NLS Ple35 CLDN5 pEMS1506 intron-lacZ/NLS Ple36...ICMT pEMS1593 intron-lacZ/NLS Ple123 ICMT pEMS1594 intron-lacZ/NLS Ple124 ICMT pEMS1595 intron-lacZ/NLS Ple125... pEMS1597 intron-lacZ/NLS Ple129 MKI67 pEMS1600 intron-lacZ/NLS Ple131 MKI67 pEMS1602 intron-lacZ/NLS ...pEMS1650 intron-lacZ/NLS Ple179 RLBP1L2 pEMS1651 intron-lacZ/NLS Ple180 RLBP1L2 pEMS1652 intron-lacZ/NLS...

  13. Brain Initiative Collection

    Type
    Collection
    ...in forebrain GABA-ergic interneurons under the control of the hDlx enhancer element Gordon Fishell 83894...in forebrain GABA-ergic interneurons under the control of the hDlx enhancer element Gordon Fishell 83895...

  14. Caltech Systemic Capsids

    Type
    Collection
    ...undergo quality control, including AAV titering by probe-based droplet digital PCR, in vitro and (when possible...#103005) . Note on PHP.eB tropism and specific mouse lines: Enhanced CNS tropism exhibited by the PHP.eB...Description Category PI Controls 28306 pAAV-FLEX-tdTomato CAG tdTomato, Cre-dependent Control Boyden 37825 pAAV-CAG-GFP...pAAV-CAG-GFP CAG GFP Control Boyden 50469 pAAV-CaMKIIa-EGFP CamKIIa EGFP Control Roth 59462 pAAV-CAG-tdTomato...pAAV-CAG-tdTomato CAG tdTomato Control Boyden 83900 pAAV-mDlx-GFP-Fishell-1 Dlx GFP Control Fishell 99130 pAAV-mDlx-NLS-mRuby2...-NLS-mRuby2 Dlx mRuby2 Control Gradinaru 104055 pAAV-CAG-eYFP CAG EYFP Control Gradinaru 104061 CAG-NLS-GFP...-GFP CAG NLS-GFP Control Gradinaru 105530 pAAV.CMV.PI.EGFP.WPRE.bGH CMV EGFP Control Wilson 105547 pENN.AAV.EF1a.eGFP.WPRE.rBG...

  15. Penn Vector Core Partnership with Addgene

    Type
    Collection
    ...pAAV-Ef1a-DIO EYFP Control Karl Deisseroth AV-1-ALL854 51502-AAV1 AAV pCAG-FLEX-EGFP-WPRE Control Hongkui Zeng....LacZ.bGH Control James M. Wilson AV-1-PV0105 105532-AAV1 pAAV.CMV.ffLuciferase.SV40 Control James M. ...eGFP.WPRE.rBG Control James M. Wilson AV-1-PV1963 105542-AAV1 pENN.AAV.CB7.CI.eGFP.WPRE.rBG Control James M....pENN.AAV.CMVs.TurboRFP.WPRE.RBG Control James M. Wilson AV-1-PV2642 105552-AAV1 pENN.AAV.hSyn.TurboRFP.WPRE.RBG Control James M...eYFP.WPRE.hGH Control Karl Deisseroth AV-1-PV3248 105557-AAV1 pENN.AAV.CB7.CI.mCerulean.WPRE.RBG Control James....SV40 Control Loren Looger AV-1-PV3661 98927-AAV1 pENN.AAV.CAG.Flex.GFPsm_myc.WPRE.SV40 Control Loren ....EGFP.WPRE.bGH Control James M. Wilson AV-2-27056 27056-AAV2 pAAV-Ef1a-DIO EYFP Control Karl Deisseroth...

  16. Adeno-associated virus (AAV) Plasmids

    Type
    Collection
    ...81070 rAAV2-retro helper AAV retrograde AAV packaging plasmid, expressing Rep2 and retrograde capsid Karpova...Baculovirus-mediated rAAV production: Expresses bicistronic AAV-2 rep gene and polycistronic AAV-1 cap gene in Sf9 insect...Baculovirus-mediated rAAV production: Expresses bicistronic AAV-2 rep gene and polycistronic AAV-2 cap gene in Sf9 insect...Baculovirus-mediated rAAV production: Expresses bicistronic AAV-2 rep gene and polycistronic AAV-6 cap gene in Sf9 insect...Baculovirus-mediated rAAV production: Expresses bicistronic AAV-2 rep gene and polycistronic AAV-8 cap gene in Sf9 insect...non-standard), expressing Rep2 and PHP.eB Cap controlled by tTA-TRE amplification system Gradinaru 103006...(non-standard), expressing Rep2 and PHP.S Cap controlled by tTA-TRE amplification system Gradinaru 127847...

  17. Plasmids for Stem Cell Research

    Type
    Collection
    ...Single polycistronic lentiviral vector for the expression of human Oct4, Sox2, and Klf4 Polycistronic lentiviral...Minicircle Human Non-integrating mini-intronic plasmid (MIP) polycistronic expression of human Oct4, Klf4, ...MMLV-derived Retrovirus Human Yamanaka factors for generating human iPS cells; retroviral expression of...MMLV-derived Retrovirus Mouse Original set of Yamanaka factors for generating mouse iPS cells; retroviral expression...30. Zhou Astrocytes Dopaminergic Neurons Lentiviral Human Efficient conversion of astrocytes to functional...Fibroblasts Astrocytes Lentiviral Mouse Direct conversion of fibroblasts into functional astrocytes by defined...27(5):1042-9. Townes Lentivirus Human Single polycistronic lentiviral vector for the expression of human...

  18. Cre-lox system

    Type
    Collection
    ...Cepko CL. 2007. Controlled Expression of Transgenes Introduced by In Vivo Electroporation. Proc Natl Acad...pLOX-CW-CRE Cre CMV Lentiviral Trono 12265 pHR-CMV-nlsCRE Cre CMV Lentiviral Trono 12493 p259 pCMV-CRE-M (BglII...Tamoxifen inducible Retroviral Jacks 24064 MSCV-Cre Cre and EGFP coexpression Retroviral Littman 24333 ins-cre_cryaa-Venus...inducible PGK Retroviral Lowe 33342 LGmCreER (non-self deleting) Cre-ERT2 PGK Retroviral Lowe 33344 pLEW100cre-EP1...pMSCVpuro-Cre Cre 5'LTR Retroviral Ge 34565 pMSCVhygro-Cre Cre and HygroR coexpression Retroviral Ge 34568 WZLneo-Cre...GFP coexpression CAG Retroviral Gage 49054 CAG-GFP/cre Cre-GFP fusion CAG Retroviral Gage 49056 AAV-GFP/...AAV Deisseroth 59701 pRetroQ-Cre-ERT2 Cre-ERT2 -Tamoxifen inducible cmvIE Retroviral Youle 59720 Nanog-CreER...

  19. New England Biolabs Cell-Imaging Plasmid Collection

    Type
    Collection
    ...pSNAPf-Cox8A Control Plasmid SNAP-tag Control for mitochondrial localization 101124 pSNAPf-H2B Control Plasmid...SNAP-tag Control for nuclear localization 101123 pSNAPf-ADRβ2 Control Plasmid SNAP-tag Control for cell...pCLIPf-H2B Control Plasmid CLIP-tag Control for nuclear localization 101130 pCLIPf-Cox8A Control Plasmid ...CLIP-tag Control for mitochondrial localization 101125 pCLIPf-NK1R Control Plasmid CLIP-tag Control for cell...pMCP-GPI Control Plasmid ACP/MCP-tag Control for cell surface localization 101131 pACP-GPI Control Plasmid...-tag Control for cell surface localization 101128 pACP-ADRβ2 Control Plasmid ACP/MCP-tag Control for cell...mammalian expression 101133 pSNAP-CaaX Control Plasmid SNAP-tag Control for endomembrane (CaaX box) localization...

  20. Antibody Guide

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    Collection
    ...negative control. The negative control may be a no-antibody control in which a sample is processed without ...standard curve can serve as the positive control); and a negative control, a sample known not to contain your...interest. The negative control should be biological (no protein expressed), which controls for your antibody...fragmentation of the DNA. Controls for capture methods Capture methods should use a positive control and a negative...addition to the control used in standard IP, ChIP should additionally use a beads-only control which runs ...the machine. Controls for cell sorting methods Flow cytometry requires a negative control, a small subset...animal’s immune system. The antigen of interest is introduced to the animal, which then mounts an immune response...
Showing: 41 - 60 of 912 results