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Are the NxSeq Single Indexing Kits (Item. No. 15100-1, 15200-1) compatible with the NxSeq UltraLow DNA Library Kit, 96 reactions (Item. No. 15096-2)?

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

Yes. If you are doing lower multiplexing and only want/need single indexes, you can absolutely combine these kits. You will reduce the cost of your library prep by buying your library prep reagents in bulk.

What kinds of samples can I use with the MasterPure Yeast Kits?

MPY03100, MPY80200

The kits can be used with a variety of yeast species, including Candida, Saccharomyces, Pichia, and Schizosaccharomyces. Additionally, they are also suitable for use with filamentous fungi, such as Aspergillus and Penicillium.

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 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).

Can I use other adaptors and/or indexing primers with your library prep kit?

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

Yes. After the End-repair/A-tailing step, the DNA fragments are A-tailed (single 3’ A overhang). These fragments are compatible with any adaptors that have a compatible T-overhang. Barcoded adaptors (e.g. TruSeq adaptors) can be ligated to the A-tailed fragments and the resulting library can be amplified with standard P5/P7 primers. However, our Universal Adaptor is optimised to produce the highest ligation efficiency with limited amounts of input DNA, and the use of other adaptors may decrease ligation efficiency and produce lower quality libraries. Also note that other PCR indexing primers may not be compatible with our Universal Adaptor, and conversely, other adaptors may not be compatible with our NxSeq Indexing Primer Sets. For optimal results, we recommend using our NxSeq Single or Dual Indexing Kits with the NxSeq UltraLow DNA Library Kit, 12 or 96 reactions.

Are the Single Index sequences the same as the Illumina TruSeq LT Adaptor sequences?

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

Yes. When setting up a sample sheet in Illumina Experiment Manager, select TruSeq LT from the drop down menu. Use A001-A027 when entering sample information, since these indexes correspond to the same numbers used for the indexes used in the NxSeq Single Indexing Kits. E.g. Illumina A001 and A027 = Biosearch Technologies 1 and 27, respectively.

Are the Dual Index sequences the same as the Illumina TruSeq HT Primer Index sequences?

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

Yes. When setting up a sample sheet in Illumina Experiment Manager, select TruSeq HT from the drop down menu. Use D701-712; D501-508 when entering sample information, since these indexes correspond to the same numbers in the NxSeq Dual Indexing Kits (E.g. Biosearch Technologies 501 = Illumina D501, Biosearch Technologies 701 = Illumina D701, etc.).

When should I use a NxSeq Single Indexing Kit vs the NxSeq Dual Indexing Kit?

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

The NxSeq Single Indexing Kits can be used for construction and pooling of 1 to 24 libraries. The NxSeq Dual Indexing Kit can be used for construction and pooling of 1 to 96 libraries. Remember that you will need to use the correct sample sheet that corresponds to either single or dual indexed libraries.

Can I pool libraries prepared with a both single- and dual-indexed libraries?

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

For most Illumina sequencing platforms, a mixture of single-and dual-indexed adapters is supported with minor modifications to the Illumina sample sheet. Note that dual indexing is not compatible with parallel Illumina platforms such as HiSeq X Ten, and a single/dual index mixture cannot be used with these platforms. Please contact our Tech Support team techsupport@lgcgroup.com for additional information regarding pooling single- and dual-indexed libraries.