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Successful CRISPR genome editing in hard-to-transfect cells (i.e., Jurkat cells)

Use the conditions presented here for Clone E6-1 Jurkat cells as a starting point for optimization of CRISPR reagent delivery in cell types requiring electroporation.

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Assessing formulas that calculate Tm—do nearest-neighbor parameters matter?

Should programs that predict Tm include nearest neighbor parameters? Learn which factors are critical for Tm calculation accuracy, and which online software takes them into account.

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Designing sgRNAs for CRISPR/Cas9 experiments

Learn how CRISPR crRNA and tracrRNA sequences can be combined into a single sgRNA for simplified use in CRISPR RNA-guided genome editing.

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Could your PCR be affected by contamination?

Learn how to prevent false amplification from DNA contamination.

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Troubleshooting Polyacrylamide Gel Electrophoresis (PAGE)

Our gel electrophoresis team provides an extensive troubleshooting guide for the PAGE issues they have come across during their work.

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qPCR Probes—selecting the best reporter dye and quencher

Recommendations for choosing dyes and quenchers, taking into account instrument compatibility and multiplex probe applications.

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NGS Target Capture Custom Panels—Determining Probe Number and Cost

The experimental design options that will determine the number (and cost) of probes required for an NGS target capture panel.

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Do your qPCR assays come with sequence information? They should. Here Is why.

qPCR assays (primer & probe sets) from other suppliers are often provided without sequence information. IDT always gives you the sequences to the oligos you order. And that can be very important. Read why.

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Recommended dye combinations for multiplex qPCR

Recommendations for selecting dyes for multiplex qPCR that minimize background and avoid overlap of fluorescent signals. Included is a table of compatible dyes for multiplexing on common qPCR instruments and a list of suggested quenchers.

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My Oligos Have Arrived: Now What?

Recommendations for resuspension and storage of newly received oligonucleotides.

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Designing PCR Primers and Probes

General guidelines for designing primers and probes and for choosing target locations for PCR amplification.

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Calculations: Converting from nanograms to copy number

Calculation often used when creating a qPCR standard curve. Link to free, online tool that will do it for you.

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GAPDH, a Good Reference Sequence?

Housekeeping genes commonly used as internal controls, often may not be the best choices due to lesser known functions and the presence of pseudogenes, some of which, in the case of GAPDH---used here as an example--may be expressed.

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Easily-designed standard curves for qPCR

An easy way to combine control templates/multiple targets onto a single construct and the advantages that can provide for PCR experiments.

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Which type of oligo purification should I choose?

Recommendations for oligonucleotide purification based on oligo length, application, yield required, and presence of modifications.

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Oligo Quantification—Getting it Right

Why a supplier's yield readings can differ from what the researcher calculates after resuspension, and the importance of using the [right] molar extinction coefficient in calculations of oligonucleotide concentration.

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Deletions During Cloning

Explanations for why sequenced clones sometimes show deletions and some suggestions for mitigating this result.

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Annealing oligonucleotides

Tips on making double-stranded DNA from single-stranded, complementary oligonucleotides.

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Calculation tips for resuspending and diluting nucleic acids

Easy guidelines for making a 100 µM solution; calculating nanomoles, micrograms, copy number, and concentration; and determining concentration equivalencies

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Sample preparation for successful qPCR

Considerations for qPCR sample preparation for obtaining accurate and consistent results.

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Steps for a successful qPCR experiment

Considerations for 5′ nuclease assay design and experimental setup to help you obtain accurate and consistent results.

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Running agarose and polyacrylamide gels

One of the most widely used tools in molecular biology, electrophoresis provides a simple, low-cost way to separate nucleic acids based on size for quantification and purification. Get some tips on running your gels.

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Tips for Using BLAST to Locate PCR Primers

Need a quick way to find the location of primers within a gene or the expected size of the resultant PCR product? In this tip we show you how to get this information using BLAST.

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