Cell Analysis Protocols

Experiment Protocol

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Immunoprecipitation Protocol Using Anti-GFP Antibodies

Experiment Protocol

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ActinRed™ 555 ReadyProbes® Reagent Protocol

Experiment Protocol

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NucGreen® Dead 488 ReadyProbes® Reagent Protocol for Fixed Cells

Experiment Protocol

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NucGreen® Dead 488 ReadyProbes® Reagent

Experiment Protocol

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CellTrace™ CFSE Cell Proliferation Kit Protocol

Experiment Protocol

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LIVE/DEAD® Fixable Dead Cell Stains

Experiment Protocol

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CellTrace™ Violet Cell Proliferation Kit Protocol

Experiment Protocol

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BrdU Labeling and Detection Protocol

Experiment Protocol

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PrestoBlue® Cell Viability Reagent for Microplates Protocol

Experiment Protocol

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LIVE/DEAD® Violet Viability/Vitality Kit Protocol

Experiment Protocol

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How can I send .sds and .sdm data files to the Real Time PCR Technical Support team?

Product FAQ

Answer

These files are often large (>5 MB) and do not always transfer well through standard email. Please visit “Contact Us” on our website and reach out to your local Technical Support team first. They can then advise you on other methods we can use to share large files with you, such as .ftp or similar programs. Please be prepared to provide us with a detailed summary of your experiment, including information such as your instrument model number, assay ID, template input, master mix used, and thermal profile.

Answer Id: E2461

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6d61bc22da834feeac4224b0973dbf1e_FAQ

Can I use probes with a Black Hole Quencher (BHQ) on Applied Biosystems® Real-Time PCR Systems?

Product FAQ

Answer

Yes, you can use probes with a BHQ on Applied Biosystems® Real-Time PCR Systems. However, you will need to set the quencher to NONE. Here is an example of a quencher setting based on the SDS software for the Applied Biosystems® 7500 Real-Time PCR System:

(1) After you create a new file, please click on TOOLS in the menu bar and choose DETECTOR MANAGER.
(2) Click FILE button on the lower left corner of the DETECTOR MANAGER window and choose NEW.
(3) In the NEW DETECTOR window, choose (NONE) in the dropdown list of QUENCHER DYE.

Answer Id: E2462

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7c1188f54e08d22268f338770150ec02_FAQ

What is the concentration of primers in the primer-limited TaqMan® Endogenous Control Assays and what function does it have in my reaction?

Product FAQ

Answer

The final primer concentration for the primer-limited Endogenous Controls is 150 nM for the forward primer and 150 nM the reverse primer. Usually, the endogenous control genes are abundantly expressed. The limited primer concentration will force the PCR of the endogenous control to plateau early, so that it leaves enough dNTPs and enzyme for lower expressed target genes to be amplified.

Answer Id: E2463

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bdf1443e71fa16257cc391ab64e0336b_FAQ

Can I check the end product of a TaqMan® Gene Expression Assay by running an agarose gel?

Product FAQ

Answer

Yes, you can check the end product on an agarose gel. If you use the TaqMan® Universal PCR Master Mix (including AmpErase® UNG, Cat. No. 4304437), we recommend running the gel immediately after the PCR, since the trace amount of AmpErase® UNG may digest the end product that contains double-stranded DNA with dUTP incorporated. The average length of amplicon ranges from 50-150 bp. The exact amplicon length is indicated on the detailed page of each assay on our website. You can get to that page by clicking on the Assay ID; the amplicon information is listed in the section "Assay Details".

Answer Id: E2464

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