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We narrowed to 1,045 results for: STI

Showing: 41 - 60 of 1045 results

  1. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ...gel from the plastic wrapper and rinse with deionized water. Gently remove the white sticker on the bottom...Rocking platform Fume hood Metal spatula Razor blade Plastic tray Gel imaging system ImageJ or similar software...Section 2: Staining the Gel Place the gel in a plastic tray with 100 mL of deionized water. Rinse gel ...the standard curve is y = 14725x + 3748.4, then substitute 18179.442 for y and solve for x. 18179.442 = ...

  2. Western Blot

    Type
    Protocol
    ...follows: Remove the gel from the plastic packaging. Remove tape and plastic comb. Rinse the wells with deionized...well microtiter plate Chemiluminescence substrate Plastic wrap Primary antibody Secondary antibody Deionized... white separator. Keep the Bottom Stack in the plastic tray. Place the Bottom Stack on the blotting surface...for 5 min at RT . Cover the membrane with clear plastic and use a gel imager with chemiluminescence detection...

  3. Protocols for Molecular Biology, Plasmid Cloning, and Viral Preps

    Type
    Protocol
    ...molecular biology, plasmid cloning, and titering and testing your viral preparations. ...free to email us at [email protected] with any questions. Intro to the Lab Bench Basic Molecular Biology...Amplification Amplify CRISPR pooled-plasmid libraries Diagnostic Restriction Digest Verify a plasmid using restriction... mouse genes. Virus Protocols for titering and testing your virus preparations. Name Description Link ...

  4. Gibson Assembly Protocol

    Type
    Protocol
    ... Gibson and colleagues at the J. Craig Venter Institute developed a novel method for the easy assembly...the assembled parts. Tips and Troubleshooting “Stitching” fragments together using oligos When you need...sequence between two PCR products, one method is to “stitch” together several oligos. This technique is especially...Pro-Tip Please note that the way to design the “stitching” primers and the amounts of primers to include...

  5. Lentivirus Production

    Type
    Protocol
    ...controller Hazardous waste container pH meter Stir plate Magnetic Stir Bar Reagents DMEM high glucose, Corning... of powder in 100 mL of deionized water. While stirring, slowly add hydrochloric acid until the solution...any packaging cells that were collected during harvesting. Filter supernatant through a 0.45 μm PES filter...

  6. Using a Light Microscope Protocol

    Type
    Protocol
    ...allows you to distinguish objects within the sample. You can increase contrast by a) adjusting the aperture.... You can take entire courses on microscopy and still have more to learn, so, for this protocol, we'll...image larger) and resolution (the ability to distinguish between two discrete objects). Figure 1 depicts...

  7. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...details on doing diagnostic digests and how to interpret them please see the Diagnostic Digest page. Purifying... how to perform gel electrophoresis. Equipment Casting tray Well combs Voltage source Gel box UV light...the solution heats up.). Caution HOT! Be careful stirring, eruptive boiling can occur. Pro-Tip It is a good...

  8. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...mammalian antibiotic resistance (e.g., puromycin, blasticidin), which enables selection of a stable cell culture...untransduced cells, resulting in a more homogenous (but still polyclonal) cell population. Depending on the transducibility...Appropriate antibiotic for selection (e.g. puromycin, blasticidin) Reagent Preparation DMEM Complete: 10% v/v FBS...lentiCas9-Blast and then selected with 1 µg/mL blasticidin for 9 days. Single cells were then plated in...

  9. CRISPR Library Amplification

    Type
    Protocol
    ...later. At a minimum we recommend the use of a diagnostic digest and high-throughput next generation sequencing...plasmid backbone a single time and visualize that digestion on an agarose gel ( see restriction digest protocol... Transform, recover, set up overnight growth (Estimated time 2-3 hours) Transformation should be performed...limited. Day 2: Harvest cells and purify DNA (Estimated time 3-4 hours) Cells should be harvested first...

  10. Plasmid Modification by Annealed Oligo Cloning (with Protocols)

    Type
    Protocol
    ...desired restriction sites and add them to your existing vector. It is a couple days of work that will ...the overhangs generated by restriction digest of existing sites in the original vector. Designing oligos...bases to complement the overhangs generated when digesting the vector with EcoRI and SalI (see diagram )....

  11. Water Bath Protocol

    Type
    Protocol
    ...Equipment Water Bath 10% bleach or 70% ethanol Distilled water Disinfectant Thermometer Water bath weights...the edge of the slide. Fill the water bath with distilled water, not tap water, to prevent salts from accumulating...water in it from previous uses. However, you will still need to regularly clean out the water bath even ...

  12. Fluorescence Titering Assay

    Type
    Protocol
    ...percentage of fluorescent cells, these cells can still be used in downstream applications. Safety Warnings...ensure that you are in compliance with your institution’s biosafety regulations. Workflow Timeline Day...with multiple integration events leading to underestimation of the true titer. Calculate the transduction...

  13. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ...genome-containing virions. Phenol red is added to clearly distinguish the steps. This method will help enrich the prep...ultracentrifugation. The steps are clearly distinguishable thanks to the use of phenol red. Image adapted...cartoon indicating the position of the needle for harvesting of the purified AAV using option #1. Right image...

  14. Pouring LB Agar Plates

    Type
    Protocol
    ... bath Pipetman Reagents 37 g pre-mixed powder consisting of: 5.0 g yeast extract 10.0 g peptone from casein...mL of agar and on which you can individually distinguish a maximum of ~100 colonies Pro-Tip If you will...After overnight drying, we place the plates in a plastic bag with an absorbent material to reduce condensation...

  15. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...lentiCas9-Blast 1 and then selected with 1 μg/mL blasticidin for 9 days. Single cells were then plated in ...transduced (MOI = 37) and selected with 1 μg/mL blasticidin for 9 days. Single cells from a stable cell pool...then expanded for an additional 3 weeks under blasticidin selection. Anti-Cas9 Western blotting was performed...

  16. Lentivirus ddPCR Titration

    Type
    Protocol
    ...ensure that you are in compliance with your institution’s biosafety regulations. Last Update: July 7, ... µg/mL. Mix well and incubate 72 h. Benzonase Digestion and Cell Harvest Gently aspirate media from the... When analyzing data there should be a clear distinction between negative droplets (black) and positive...

  17. Biosensor AAV Preps

    Type
    Collection
    ...iAChSnFR none Constitutive 1 Looger 137955 pAAV.CAG.iAChSnFR CAG iAChSnFR none Constitutive 1, 9 Looger ... none Constitutive 1, 5, rg* Looger 176753 AAV-mDlx-jGCaMP8f-WPRE Dlx jGCaMP8f none Constitutive 1, 9,...jGCaMP8f none Constitutive 1, 9 Looger 179254 AAV-CAG-jGCaMP8f-WPRE CAG jGCaMP8f none Constitutive 1, 9, rg...ribo-jGCaMP8s none Constitutive 1 Fyhn 169256 AAV-hSyn-Soma-jGCaMP8s Syn soma-jGCaMP8s none Constitutive 1, 9, rg...none Constitutive 1, 5, 9, rg* Looger 176755 AAV-mDlx-jGCaMP8s-WPRE Dlx jGCaMP8s none Constitutive 1, 9...jGCaMP8s none Constitutive 1, 9 Looger 179256 AAV-CAG-jGCaMP8s-WPRE CAG jGCaMP8s none Constitutive 1, 9, rg...ribo-jGCaMP8m none Constitutive 1 Fyhn 169257 AAV-hSyn-Soma-jGCaMP8m Syn soma-jGCaMP8m none Constitutive 1, 9, rg...

  18. Control AAV Preps

    Type
    Collection
    ...gfaABC1D tdTomato Constitutive 5 Baljit Khakh 50465 pAAV-hSyn-EGFP hSyn EGFP Constitutive 1, 2, 5, 8, 9, ...tdTomato Constitutive 5 Hongkui Zeng 58909 pAAV-GFAP104-mCherry GFAP104 mCherry Constitutive 5 Edward ...PI.eGFP.WPRE.bGH GFA104 EGFP Constitutive 5 Philip Haydon 104055 pAAV-CAG-eYFP CAG EYFP Constitutive 2, 5, rg*, PHP....-GFP Constitutive PHPeB Viviana Gradinaru 105530 pAAV.CMV.PI.EGFP.WPRE.bGH CMV EGFP Constitutive 1, 2,...ffLuciferase Constitutive 8 James M. Wilson 105534 pAAV.TBG.PI.LacZ.bGH TBG LacZ Constitutive 8 James M....TBG None Constitutive 8 James M. Wilson 105539 pAAV.hSyn.eGFP.WPRE.bGH Syn EGFP Constitutive 1 James M...CamKII EGFP Constitutive 1, 5 James M. Wilson 105535 pAAV.TBG.PI.eGFP.WPRE.bGH TBG EGFP Constitutive 8 James...

  19. Optogenetics AAV Preps

    Type
    Collection
    ...mScarlet Constitutive 9 Christopher Harvey 59170 pAAV-Syn-Chronos-GFP Syn Chronos GFP Constitutive 1, 5, ...ChETA EYFP Constitutive 9 Karl Deisseroth 135633 pAAV-S5E2-C1V1-eYFP E2 C1V1 EYFP Constitutive 1, 5, 9, ...mCherry Constitutive 8 Roger Tsien 99039 pAAV-CamKII-ArchT-GFP (PV2527) CaMKII ArchT GFP Constitutive 1, 5...EYFP Constitutive 2, 5 Karl Deisseroth 137151 pAAV-nEF-NpHR3.3-EYFP nEF NpHR 3.3 EYFP Constitutive 8 Karl...dependent Flp dependent Cre and Flp dependent Constitutive Serotype AAV1 AAV2 AAV5 AAV8 AAV9 AAVrg PHPeB...PI 20071 pACAGW-ChR2-Venus-AAV CAG ChR2 Venus Constitutive 1, 9 Karel Svoboda 83898 pAAV-mDlx-ChR2-mCherry-Fishell...mCherry-Fishell-3 Dlx ChR2 mCherry Constitutive 1, 9, rg* Gordon Fishell 135634 pAAV-S5E2-ChR2-mCherry nEF...

  20. Fluorescent Protein Guide: Biosensors

    Type
    Collection
    ...activity (GCaMP6f/m/s) (Constitutive or Cre-dependent) Cell-specific restoration of stimulus preference after... experimental design, biosensors can enable investigation of a signaling pathway or measurement of a biomolecule...a biosensor in a novel context or modifying an existing construct, as fluorescence properties can be affected...Fast Genetically Encoded Calcium Indicators (Constitutive or Cre-dependent) jGCaMP8 Fast Genetically Encoded...High-performance GFP-based calcium indicators (Constitutive or Cre-dependent) High-performance calcium sensors...calcium sensors for imaging neural activity (Constitutive or Cre-dependent) Ultrasensitive fluorescent...regulatory element Viral manipulation of functionally distinct interneurons in mice, non-human primates and humans...
Showing: 41 - 60 of 1045 results