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What is the best way to quantify DNA or RNA isolated using the MasterPure Complete Kit?

MC89010, MC85200

Spectrophotometric methods (e.g., A260) to quantify DNA or RNA, although in common use, may overestimate the concentration. The best method to quantify DNA is by fluorometry using a DNA-specific dye, such as Hoechst 332581 (bisbenzimide), or PicoGreen® dye (Thermo Fisher Scientific). These dyes bind specifically to double-stranded DNA and not to nucleotides, single-stranded DNA, or RNA. To quantify RNA, we recommend using a Qubit™ fluorometer (Thermo Fisher Scientific) or a 2100 BioAnalyzer® instrument (Agilent).

Do I need to perform the optional RNase treatment step in the MasterPure Yeast DNA Purification Kit protocol?

MPY03100, MPY80200

We recommend treating the cell lysate with RNase A (supplied in the kit) if the presence of small amounts of RNA will interfere with downstream applications, such as some PCR applications or sequencing library preparations. However, for applications such as restriction enzyme digestion and cloning, or PCR with primers specific to genomic DNA sequences, RNase treatment is not necessary.

Do I need to perform the optional DNase treatment step in the MasterPure Yeast RNA Purification Kit protocol?

MPY03100, MPY80200

The MasterPure Yeast RNA Purification Kit protocol recovers DNA with much lower efficiency than RNA after cell lysis, so removal of contaminating DNA may not be necessary for many applications. For applications such as RT-PCR with gene-specific primers, we recommend following the DNase treatment protocol.

Is BHQ Probe Master Mix stable at room temperature?

BHQ-001

BHQ Probe Master Mix is an incredibly stable PCR/qPCR Master Mix, which shows little-to-no reduction in performance when left at room temperature, even when combined with all other PCR-reaction components. We have demonstrated Cq > 1.0 when assembled qPCR reactions were left at room temperature for 16 hours, in both singleplex and multiplex applications.

To maintain high performance, it is recommended to store BHQ Probe Master Mix at -20 °C, and aliquot it into nuclease-free, light-protected tubes to minimise the number of freeze-thaw cycles. BHQ Probe Master Mix is also stable at +4 °C for 1 week.

I want to amplify a template that is ~20 kb. Should I use the FailSafe PCR System or the MasterAmp™ Extra-Long PCR Kit?

FS99100, FS99250, FS9901K, PCRAMP-018

The FailSafe PCR System can amplify templates ~20 kb. For templates >10 kb, we recommend using 2.5 units of the FailSafe PCR Enzyme Mix (for templates <10 kb, use 1.25 units) in 50 µL reactions.

What is the best way to quantify DNA purified using the MasterPure Gram Positive Kit?

MGP04100, MB711400

Spectrophotometric methods (e.g., A260) to quantify DNA or RNA, although in common use, may overestimate the concentration. The best method to quantify DNA is by fluorometry using a DNA-specific dye, such as Hoechst 33258 (bisbenzimide), or PicoGreen® dye (Thermo Fisher Scientific). These dyes bind specifically to double-stranded DNA and not to nucleotides, single-stranded DNA, or RNA.

What is the best way to quantify DNA or RNA isolated using the MasterPure Yeast Kits?

MPY03100, MPY80200

Spectrophotometric methods (e.g., A260) to quantify DNA or RNA, although in common use, may overestimate the concentration. The best method to quantify DNA is by fluorometry using a DNA-specific dye, such as Hoechst 33258 (bisbenzimide), or PicoGreen® dye (Thermo Fisher Scientific). These dyes bind specifically to double-stranded DNA and not to nucleotides, single-stranded DNA, or RNA. To quantify RNA, we recommend using a Qubit™ fluorometer (Thermo Fisher Scientific) or a 2100 BioAnalyzer® instrument (Agilent).

What is the best way to quantify DNA purified using the MasterPure Gram Positive Kit?

MGP04100, MB711400

Spectrophotometric methods (e.g., A260) to quantify DNA or RNA, although in common use, may overestimate the concentration. The best method to quantify DNA is by fluorometry using a DNA-specific dye, such as Hoechst 332581 (bisbenzimide), or PicoGreen® dye (Thermo Fisher Scientific). These dyes bind specifically to double-stranded DNA and not to nucleotides, single-stranded DNA, or RNA.

What is the best way to quantify DNA or RNA isolated using the MasterPure Complete Kit?

NAEXPU-001, MB711400

Spectrophotometric methods (e.g., A260) to quantify DNA or RNA, although in common use, may overestimate the concentration. The best method to quantify DNA is by fluorometry using a DNA-specific dye, such as Hoechst 33258 (bisbenzimide), or PicoGreen® dye (Thermo Fisher Scientific). These dyes bind specifically to double-stranded DNA and not to nucleotides, single-stranded DNA, or RNA. To quantify RNA, we recommend using a Qubit™ fluorometer (Thermo Fisher Scientific) or a 2100 BioAnalyzer® instrument (Agilent).

Can I perform both singleplex and multiplex assays using the RapiDxFire Lyo-Flex 1-Step RT-qPCR 5X Master Mix?

PCRAMP-021

Yes.