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We narrowed to 286 results for: lum

Showing: 241 - 260 of 286 results

  1. Chemogenetics Guide

    Type
    Guide
    ...collection of PSAM plasmids ! LMOs Luminopsins (LMOs), or luminescent opsins, are opto-chemogenetic fusion...laser or LED illumination), making LMOs a versatile option. Coupling brighter bioluminescent modules, such...Gross RE. (2015). Inhibitory luminopsins: genetically-encoded bioluminescent opsins for versatile, scalable...systems in our Optogenetics Guide . Figure 4: Luminopsins can be activated through chemogenetic or optogenetic...Augustine GJ, Yin HH, Hochgeschwender U (2016). Luminopsins integrate opto- and chemogenetics by using physical... a single molecule driven by FRET-modified bioluminescence. Neurophotonics. 11:021005. PMID: 38450294 ...Shaner NC, Hochgeschwender U (2024). Engineering luminopsins with improved coupling efficiencies. Neurophotonics...

  3. CRISPR Guide

    Type
    Collection
    ...with higher fidelity than SpCas9. FrCas9 (from Faecalibaculum rodentium ) has increased fidelity, and recognizes...

  4. 15 Hot Plasmids from 2017

    Type
    Blog Post
    ...determined by the organized smooth endoplasmic reticulum (OSER) assay. A major advantage of mCyRFP1 is ...

  6. Optogenetics Guide

    Type
    Guide
    ...sequestered Lumitoxin Peptide toxin for blocking K + (Kv) channels fused to LOV2 photoswitch. Illumination causes...expression of the opsin and the area that is being illuminated. There are several different ways to control ...2006. Next-generation optical technologies for illuminating genetically targeted brain circuits. J Neurosci...

  7. Molecular Biology Reference

    Type
    Guide
    ...acquire NGS but one of the most commonly used is the Illumina NGS platform. This is the platform used by Addgene...sequencing partner, seqWell . The actual process of Illumina NGS is not that different from Sanger sequencing...modified fluorescently tagged nucleotides. During illumina NGS, a long piece of DNA is first fragmented into...regular base and thus do not halt the reaction. Illumina NGS, therefore does not require any “normal” bases...

  8. Antibody Guide

    Type
    Guide
    ...performs a chemiluminescent reaction in the presence of several different substrates. The luminescence produced... protein extract by centrifugation or affinity columns. Mix samples with a primary antibody, which is ...

  9. Modular Cloning Guide

    Type
    Guide
    ...variety of applications. EcoFlex MoClo Extension, Volume I Bacterial Expression Mikhail Shapiro 96 cross-compatible...expression tuning in E. coli . CIDAR MoClo Extension, Volume I Bacterial Expression Richard Murray 110 plasmids...

  10. CRISPR Guide

    Type
    Guide
    ...with higher fidelity than SpCas9. FrCas9 (from Faecalibaculum rodentium ) has increased fidelity, and recognizes...

  11. Affinity Purification of Recombinant Antibodies with Protein A or Protein G

    Type
    Protocol
    ...21028 Protein G GraviTrap columns, Cytiva 28-9852-55 or rProtein A GraviTrap columns, Cytiva 28-9852-54 LabMate...Desalting Columns, 7 kDa MWCO, 10 mL, Thermo Fisher 89893 Amicon Ultra-15 30 kDa MWCO column, Millipore...Reservoirs to the top of the Gravitrap column. Pro-Tip Gravitrap columns have a finite binding capacity. If...among multiple columns. For information about the total binding capacity for your columns, refer to the...Gravitrap column to a clamp on a clamp stand. Use scissors to cut the bottom of the Gravitrap column at the...Gravitrap columns at the bottom. Add 5 mL of Pierce IgG Elution Buffer to the capped Gravitrap column and incubate...Amicon 30 kDa MWCO buffer exchange column. (Optional) Regenerate the column by washing with 25 mL of Protein...

  12. Pipetting Protocol

    Type
    Protocol
    ...liquid into the pipette tip, the volume display which shows the set volume for the the pipette, and the tip...shows the volume adjustment ring right below the plunger. This ring changes the pipette volume. Below that...with and the volume to be dispensed. The boxes that the tips come in often indicate a volume range that ...shows the pipette with the appropriate dispense volume: This image shows the P2, P10, P20, P100, P200,...P1000 as shown from left to right. Pipette Dispense Volume P2 0.2 to 2 µL P10 1 to 10 µL P20 2 to 20 µL P100...be closed to prevent contamination. Reading the Volume on the Pipette Although each pipette performs the...picture above), while 0.5 µL would read as 050. Each volume display on these pipettes will also have small ...

  13. Lentivirus ddPCR Titration

    Type
    Protocol
    ... below. Dilution Factor Virus Volume to Dilute Media Volume Total Volume 2.5 160 µL viral stock 240 µL...enough master mix for 9 samples. Component Volume 9X Volume Final Concentrations 2X ddPCR Supermix for...per well is 300,000 and the virus volume refers to the total volume the cells were transduced, which is...microcentrifuge tubes with the appropriate amount of media volume listed in the chart below. For the first virus ... well with a 1 mL pipette 5–10 times. The final volume in the well is 1.5 mL, and the final Polybrene ...nM, 250 nM Nuclease-free water 4 µL 36 µL Total Volume 16 µL 144 µL Vortex the master mix for 15 sec and... # Cells per well D = Dilution factor v = Virus volume, mL *In the experimental setup above, the cells...

  14. AAV Production in HEK293 Cells

    Type
    Protocol
    ...cell suspension (no clumps of cells). Pool cells from 2 x T-175 flasks. Adjust volume to 300 mL with DMEM...filter system, Nalgene, 565-0010 (or for larger volumes 1000 mL 0.45 µm Rapid-Flow PES Filtration Unit,...NaCl into deionized water and adjust to a final volume of 1000 mL. Stir at RT until fully dissolved. Pro-Tip...vigorously to obtain a single cell suspension (no clumps). Count cells using a hemocytometer or cell counter...million cells from the CS2 into one CS5 with a total volume of 850 mL. Return to the incubator for 24 h–36 ...Plasmid Plasmid Size (bp) DNA Concentration (μg/μL) Volume of DNA (μL) RepCap 7,265 1.00 727.6 pHelper 11,...Calculations RepCap: 0.08 μg/bp × 7,265 bp = 582.1 μg Volume Needed: 582.1 μg / 1.0 μg/μL = 582.1 μL pHelper...

  15. Handling Plasmids from Addgene - Purifying Plasmid DNA

    Type
    Protocol
    ...solution to a column that will bind the now precipitated DNA. Wash the pellet or column with 70% ethanol...your DNA solution, add 2-2.5 volumes 95% or 100% ethanol and 1/10 volume of 3 M Na-acetate (pH 4.8). Invert...-Tip Refer to appropriate DNA prep protocol for volume of bacteria to grow (low copy plasmids require ...bacteria back into suspension, but within a smaller volume of buffer that is compatible with the next solution...Resuspend the DNA pellet, or elute the DNA off of the column using water or a neutral buffer such as TE. You...Phenol-Chloroform Extraction of DNA Samples Add an equal volume of TE-saturated phenol-chloroform to the aqueous...and place it in a new microfuge tube. Add equal volume of chloroform to the recovered aqueous DNA layer...

  16. Western Blot

    Type
    Protocol
    ...cell pellet in appropriate volume of cold lysis buffer. Pro-Tips The volume of lysis buffer will vary ...Nonfat milk powder 96-well microtiter plate Chemiluminescence substrate Plastic wrap Primary antibody Secondary...the sample from the standard curve. Determine the volume of sample required to load an equivalent amount... at RT on a shaking platform. Prepare the chemiluminescence substrate by mixing 1:1 reagent A to reagent...reagent B. Gently incubate the membrane in the chemiluminescence reagent for 5 min at RT . Cover the membrane...with clear plastic and use a gel imager with chemiluminescence detection or a dark room to detect the bands...

  17. Fluorescence Titering Assay

    Type
    Protocol
    ...dilutions. Dilution Volume of Lentivirus Stock (μL) Volume of DMEM complete (μL) Volume of 10mg/mL polybrene...using either the dilution factors (method 1) or the volume of virus (method 2): Method 1 Method 2 $$T = {N...fluorescence D = Dilution Factor V T = Transduction Volume, mL For example: If 150,000 cells were transduced...Fraction of cells with fluorescence V V = Virus Volume, mL For example: If 15 µL of virus added to 150,000...

  18. AAV Purification by Iodixanol Gradient Ultracentrifugation

    Type
    Protocol
    ...bottom of the column. We recommend concentrating to a minimum of 500 µL. If the concentrate volume is less ...video below to learn setup and use an iodixanol column gradient for AAV purification. Workflow Timeline...MgCl 2 and 14.91 mg of KCl in 1× PBS in a final volume of 100 mL. Sterilize by passing through a 0.22 ...MgCl 2 , and 14.91 mg of KCl in 1× PBS in a final volume of 100 mL. Sterilize by passing through a 0.22 ...less than 500 µL, bring up the volume with formulation buffer. Use a P1000 to the bottom of the filter and...

  19. AAV Titration by qPCR Using SYBR Green Technology

    Type
    Protocol
    ...stock made in step #1): Volume of 2 x 10 9 stock or previous dilution (μL) Volume of nuclease-free water... in the table below: Dilution Series Volume of sample (uL) Volume of nuclease free water (uL) Dilution...protocol is for a 96-well plate with 20 μL reaction volume. Last Update: February 13, 2019 Estimate of time...least 10 times, and use at least half of the final volume (mix with >50 uL if your well contains 100 uL) ...

  20. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ... Determine the volume of homogenized cell solution that corresponds to 50 cells: Volume of homogenized...cell pool by trypsinization and break up any cell clumps into individual cells by passing several times ...conditioned medium, it will be more accurate if a higher volume of homogenized cell solution is transferred. Transfer...cells/mL) = 0.125 µL Because this is such a small volume, first make 1 mL of a 1:100 dilution of the homogenized...transferring 0.125 µL, now transfer 100 times that volume, which is 12.5 µL. To make the final 5 cell/mL ...
Showing: 241 - 260 of 286 results