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Ideas to Streamline Your Research

Storing oligos: 7 things you should know

Lab tips: Researchers often have questions about the stability of their oligos and how best to store them. Review these important considerations and supporting data, which were generated from an ongoing, multi-year, longitudinal stability study.

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Tips for working with gBlocks® Gene Fragments

Working with IDT custom, synthetic dsDNA fragments? Get these tips from our scientists on the best ways to resuspend, quantify, and calculate copy number of gBlocks® Gene Fragments.

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Considerations when adopting published sequences for your own qPCR assay

Lab tip: Published papers that use qPCR applications are a resource of vetted assay sequences. These can be co-opted and converted to more sensitive, double-quenched probes for use in your own experiments. Before ordering, though, go through this checklist to ensure that these sequences will work well in your assays, and consider up-to-date PrimeTime Predesigned qPCR Assays with guaranteed efficiencies of >90%.

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Genome editing tip: A CRISPR RNA annealing step can increase editing efficiency

Looking for ways to increase the genome editing activity in your CRISPR experiments? This quick test suggests that spending a little extra time to anneal CRISPR RNAs will provide improvement.

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Could your PCR be affected by contamination?

Learn how to prevent false amplification from DNA contamination, and how to address contamination when it does occur.

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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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CRISPR genome editing: 5 considerations for target site selection

Read how your genome editing experiments can be improved with 5 quick tips for target selection and with reagents from the Alt-R® CRISPR-Cas9 System.

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Genome editing: How stable is my CRISPR RNA:Cas9 RNP complex?

You can safely complex CRISPR RNAs with Cas9 in advance of your experiments and store these RNPs for future use. Store CRISPR RNPs at 4°C for up to 2 weeks, or at –80°C long-term. RNP complexes stored in this way provide the same high level of genome editing as freshly complexed RNPs.

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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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Troubleshooting polyacrylamide gel electrophoresis (PAGE)

Review this extensive troubleshooting guide for the PAGE issues our gel electrophoresis team has encountered during their work.

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qPCR Probes—selecting the best reporter dye and quencher

Read these 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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How important are those NGS metrics?

Of the many metrics used in evaluating target capture data for NGS applications, read about which ones our researchers consider important for evaluating performance of target enrichment panels.

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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?

Review these recommendations for resuspension and storage of newly received oligonucleotides.

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Designing PCR primers and probes

Many factors can influence successful PCR experiments, including primer and probe location, length, interaction and self-folding, melting temperature, annealing temperature, and GC content. Review these general recommendations for designing primers and probes and for choosing target locations for PCR amplification.

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Calculations: Converting from nanograms to copy number

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

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GAPDH, a good reference sequence?

What internal controls are you using? Housekeeping genes are frequently used as internal controls. However, they may not be the best choices due to lesser known functions and the presence of pseudogenes, some of which, as in the case of GAPDH—used here as an example—may be expressed.

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Easily designed standard curves for qPCR

Adopt this easy way to combine control templates/multiple targets onto a single construct, and get the advantages that they provide for PCR experiments.

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Which type of oligo purification should I choose?

Do your oligos need purification for your intended application? Use these recommendations based on oligo length, application, yield required, and presence of modifications to determine which oligonucleotide purification method to select.

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Oligo quantification—getting it right

Did you know that a supplier's yield readings can differ from what the researcher calculates after resuspension? Learn 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

Use this quick protocol for making double-stranded DNA from single-stranded, complementary oligonucleotides. Also review some considerations for making and using annealed oligos.

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Calculation tips for resuspending and diluting nucleic acids

Use these simple 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

Read about qPCR sample preparation and what experimental details you should consider for obtaining accurate and consistent results.

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Steps for a successful qPCR experiment

Read these recommendations for 5′ nuclease assay design and experimental setup that will 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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