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We narrowed to 395 results for: POR C

Showing: 381 - 395 of 395 results

  1. Protocol - How to Run an Agarose Gel

    Type
    Protocol
    ...attention. Let agarose solution cool down to about 50 °C (about when you can comfortably keep your hand on ... with a pipette tip. Place newly poured gel at 4 °C for 10-15 mins OR let sit at room temperature for ... set more quickly if you place the gel tray at 4 °C earlier so that it is already cold when the gel is...period of time; b) using a wider/thinner gel comb; or c) loading less DNA into the well. Another method for... be in the bottom portion of the gel, but all of the EtBr will be in the top portion and your bands will...overboil the solution, as some of the buffer will evaporate and thus alter the final percentage of agarose...

  2. Protocol - How to Ligate Plasmid DNA

    Type
    Protocol
    ...individual tubes of 5, 10 or 20μL for storage at -20°C. Whenever you need to set up ligations in the future...optimization. Incubate at room temperature for 2hr, or at 16°C overnight (following the manufacturer’s instructions...efficiency of ligation can be improved by incubation at 37°C. Proceed with bacterial transformation . Tips and ...desired insert. If you are in this situation, it is important to treat the digested vector backbone with a phosphatase...setting up the ligation reaction itself, it is important to determine the amount of cut insert and vector...being sure to increase the amount of buffer proportionally. 1μL of ligase should be sufficient for larger...

  3. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...heat-inactivated FBS and 5mL of 100X glutaGRO. Store at 4 °C. Pro-Tip Different brands and FBS lots can promote...it can be inactivated in the lab by heating to 56 °C for 30 min. Considerations Before You Start The health...dose of antibiotic, which may not be stable at 37 °C. To achieve a stable cell pool, the antibiotic selection...well plate and left undisturbed for 13 days. (a, b, c) Colonies formed by expansion of single cells for ...CI) Heat-inactivated FBS Polybrene (10 mg/mL), Millipore TR-1003-G 1X PBS pH 7.4 without calcium or magnesium...of cell death upon antibiotic selection. It is important to monitor these cells regularly and replace the... the surviving cells in the culture, so it is important to do regular media changes and maintain optimal...

  4. Gibson Assembly Protocol

    Type
    Protocol
    ... Cepko, 2020). Incubate the mix for 1 hour at 50 °C or follow manufacturer's instructions. You can purchase...Gibson DG, Smith HO, Hutchison CA, Venter JC, Merryman C. (2010). Chemical synthesis of the mouse mitochondrial....1515 (Link opens in a new window) Rabe BA, Cepko C. (2020). A Simple Enhancement for Gibson Isothermal...other. Phusion High-Fidelity DNA Polymerase - incorporates nucleotides to “fill in” the gaps in the annealed... product by restriction digest . Sequence the important regions of your final plasmid, particularly the...

  5. AAV ddPCR Titration

    Type
    Protocol
    ...parameters. Cycling Step Temperature (°C) Time (min) Ramp Rate (°C/sec) # Cycles Denaturation 95 10 2 1 ...sample: $$T = {R*C*D*1000\over V}$$ Where: T = Titer, GC/mL R = Reaction volume (20 µL) C = Copies/µL D ...Bio-Rad, 1863051 8-strip PCR tubes, Axygen, PCR-02-FCP-C ddPCR 96-well PCR plates, Bio-Rad, 12001925 48-well...place the PCR plate with the foil onto the metal support block. Place the block in the plate sealer and ... reading. When the droplet reading is complete, export the data from all wells as a CSV file and use the...

  6. Antibody Guide

    Type
    Guide
    ...stored at 4 °C for short-term storage. For long-term storage they should be stored at -20 °C in aliquots...conjugated to fluorophores should never be stored below 4 °C. Certain isotype subclasses, such as IgG3, aggregate... after thawing and therefore should be kept at 4 °C. Record and track lot numbers for all antibodies used...necessary in cases where extreme specificity is important. Their epitope specificity allows binding to a...antibodies only bind a single epitope, so it is important to choose a monoclonal antibody that will recognize...using a secondary against a tag, such as His, incorporated in the sdAb. sdAbs can have weak signals due...with conjugating reactions are possible. It is important to research your reaction and buffer before conjugating...

  7. Optogenetics Guide

    Type
    Guide
    ... PMID 22341318 Gradinaru V, Zhang F, Ramakrishnan C, Mattis J, Prakash R, Diester I, Goshen I, Thompson...20621963 Mattis J, Tye KM, Ferenczi EA, Ramakrishnan C, O'Shea DJ, Prakash R, Gunaydin LA, Hyun M, Fenno ...neural populations at distinct times. Temporal considerations. Temporal precision is key in optogenetic experiments... exposed to light of the correct wavelength, the pore opens, cations flow into the cell (yellow dots),...optical switches Sensors are genetically-encoded reporters of molecular signals; e.g., calcium indicators...specific subsets of neurons, allowing precise spatiotemporal control of these neurons by turning on and ...addition of trafficking signal from Kir2.1 and ER export signal provide improved membrane targeting 589 ...

  8. Fluorescence Titering Assay

    Type
    Protocol
    ...heat-inactivated FBS and 5 mL of 100X glutaGRO. Store at 4 °C. Pro-Tip Different brands and lots of FBS can promote...it can be inactivated in the lab by heating to 56 °C for 30 min. Considerations Before You Start The health...CI) Heat-inactivated FBS Polybrene (10mg/mL), Millipore TR-1003-G 1X PBS pH 7.4 without calcium or magnesium...

  9. CRISPR Library Amplification

    Type
    Protocol
    ...media at 37 °C (for at least 15 minutes). Prewarm 3X LB Agar + Antibiotic plates at 37 °C. Prewarm 8X ...Chill a box of 200 µL micropipette tips in a -20 °C freezer. Aliquot 3 mL SOC into each of four 14 mL ... mix. Electroporate cells (one at a time for a total of eight electroporations): Electroporator Conditions...and have 1 mL SOC per electroporation readily available for post-electroporation recovery of cells. Ensure...if the recombinant band makes up a significant proportion of the DNA pool. Last Update: August 17, 2023...limited. Equipment Table top centrifuge BioRad Electroporator (MicroPulser TM , Bio-Rad 1652100) Reagents.../or increasing the skewness of the pool. 8 electroporation cuvettes (BioRad, Micropulser, 0.1 cm ) 20 ...

  10. Colony Formation Titering Assay

    Type
    Protocol
    ...inactivated FBS and 5 mL of 100X glutaGRO. Store at 4 °C. Pro-Tip Different brands and lots of FBS can promote...it can be inactivated in the lab by heating to 56 °C for 30 min. Considerations Before You Start The health...CI) Heat-inactivated FBS Polybrene (10 mg/mL), Millipore TR-1003-G 1X PBS pH 7.4 without calcium or magnesium...

  11. Coomassie Purity Stain of Recombinant Antibodies

    Type
    Protocol
    ...standard Before Starting Warm the hot plate to 100 °C. Thaw IgG standard and prestained protein ladder on...microcentrifuge. Heat the samples for 10 min at 100 °C in a heat block. Spin the sample briefly in the microcentrifuge...IgG standards in Google Sheets. Open exported CSV file. Select import. Choose file. Format as needed. Select...that sharing the full details of our protocols supports reproducibility and accelerates science. Here,...information is solely for transparency intended to support reproducibility in science. Last Update: June 14...intensity of the protein bands to the overall lane. Import the gel image into ImageJ. Select File . Select... each peak by selecting the center of the peak. Export results as a csv file. In the results table, select...

  12. Protocol - How to Purify DNA from an Agarose Gel

    Type
    Protocol
    ...longer period of time; b) using a wider gel comb; or c) loading less DNA in the well. How do you get better...around the band as possible. To do so, it is often important to take the excised band, lay it down on the UV...sides with the razor blade. This is especially important during the DNA purification step, as many kits... fragment. The weight of the gel is directly proportional to its liquid volume and this is used to determine...manufacturer's instructions. Note: It is usually important to determine the concentration of the DNA that...

  13. Plan Your Experiment

    Type
    Guide
    ...) may be more efficacious than gRNAs containing a C nucleotide at the same position in spite of being ...promoter is typically used for gRNA May contain reporter gene (e.g. GFP) to identify and enrich positive...vector or separate transfer vectors May contain reporter gene (e.g. GFP) or selection marker to identify... to target cells through microinjection or electroporation Transient expression of CRISPR components Expression...may alter protein function when they occur in important protein domains. For gene editing experiments ... addition to off-target activity , it is also important to consider factors that maximize cleavage of ...25408407 Hashimoto, M., & Takemoto, T. (2015). Electroporation enables the efficient mRNA delivery into the...

  14. Water Bath Protocol

    Type
    Protocol
    ... refrigerators to achieve a temperature between 4°C and room temperature. This general protocol suits ...on the surfaces of the water bath as the water evaporates from the tub. Disinfectant may be added to prevent...cover on the top of the water bath to prevent evaporation and maintain the desired temperature. This also...

  15. Ligation Independent Cloning

    Type
    Protocol
    ...software to ensure a melting temperature between 50-60°C for your PCR primers. Step 2: Linearize Vector In ...provided by your polymerase manufacturer. It is very important to remove all free nucleotides from your PCR product...
Showing: 381 - 395 of 395 results