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  2. WA 0812 2782 5310 Kontraktor Pemborong Interior Rumah Minimalis Sederhana Ukuran 800x600 Daerah Ponjong Gunungkidul

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How do waste handling requirements differ between original sbeadex and sbeadex Lightning?

SBXLCK, SBXLCKSTART

Waste handling requirements do not differ between original sbeadex and sbeadex Lightning. Waste amounts and toxicity for laboratory personnel are significantly reduced with sbeadex Lightning.

My AmpliScribe reaction had excellent yields, but when I ran it on a native gel the RNA was much shorter than the original template. What could have gone wrong?

AS3107, MBTOOL-007

Single stranded in vitro transcripts are best separated using denaturing gels1. Denaturing gels allow in vitro transcripts to separate on the basis of their length rather than based on their length + secondary structure. When using native gels, sample migration may be altered by secondary structures in the transcripts.

Which FailSafe PreMix should I use for my GC-rich template? I don't want to test all of the PreMixes.

PCRAMP-020, FS99060, PCRAMP-018, PCRAMP-017

It is nearly impossible to determine, without experimentation, which FailSafe PCR PreMix will work best with a given template/primer combination. We strongly recommend using all of the PreMixes for optimisation. Without testing the complete set of FailSafe Premixes, you may not be able to determine the optimal PCR conditions for the specific amplicon that you need.

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.

Can I shorten the recommended wash buffer steps or leave a step out?

SBXMXPL, SBXMNPK, SXMXPBS, NAP44628, SBXMNPK-01, NAP41605, SXMXPBS-01, SBXMXPL-01
Depending on species and required DNA quality, the wash times in each wash step can be shortened. We recommend optimising the protocol for your specific sample type and application. Often it is sufficient to wash only once with wash buffer PN1 and once with wash buffer PN2 and omit the water wash step.

In my end-point assay, the NTC signal was the same as my target signal. Why?

The reaction ran too long and the background NTC signal was allowed to catch up to the maximum sample signal, mostly likely due to primer dimerisation or nonspecific primer annealing and extension events.

See: Why does the signal appear early in my no template control (NTC)?

In my endpoint assay, the NTC signal was the same as my target signal. Why?

The reaction ran too long and the background NTC signal was allowed to catch up to the maximum sample signal, mostly likely due to primer dimerisation or non-specific primer annealing and extension events. See: Why does the signal appear early in my no template control (NTC)?

Index hopping and misassignment of reads to the wrong sample is an issue for me. Since the NxSeq HT Dual Indexing Kit creates libraries that share the same indexes on one end or the other, is there a way that I can use these Dual Indexing Primers to create libraries with truly unique indexes at both ends, with no shared indexes across libraries?

NXGSEQ-003, NXGSEQ-004, 15300-1, NXGSEQ-001, NXGSEQ-002

Yes. Since our Dual Index Primers are provided in individual tubes, you can make and multiplex up to 8 libraries with unique dual indexes at each end. There are various combinations that will produce uniquely dual indexed libraries. For example, here are (8) unique primer combinations: (701,501), (702,502), (703,503), (704,504), (705,505), (706,506), (707,507), and (708,508). In this case, you can make and multiplex up to 8 libraries with unique dual indexes at each end. The 500 Series can also be combined with the other 700 Series Primers to more fully utilize the available primers. For example, here are two sets of four libraries that can be constructed and pooled with unique dual indexes. Set 1: (709,501), (710,502), (711,503), and (712,504). Set 2: (709,505), (710,506), (711,507), and (712,508). By using these three sets of primer combinations on separate sequencing runs, you can use all of each primer provided in the NxSeq HT Dual Indexing Kit.

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).

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.