GeneChip™ WT Terminal Labeling Kit, 30 reactions - FAQs

View additional product information for GeneChip™ WT Terminal Labeling Kit - FAQs (900671, 900670)

6 product FAQs found

How long does it take to scan an array?

It takes approximately 35 minutes to scan each Exon Array.

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Can I use the WT Sense Target Labeling Assay protocol for prokaryotic arrays?

This has not been tested at the moment; therefore, it is not recommended to use the protocol for any application other than on the Exon Arrays.

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What is the typical cRNA yield after the IVT reaction in the first cycle?

Starting with 1 µg of total RNA, following the standard protocol, ≥ 16 µg of cRNA is routinely obtained. Starting with 100 ng of total RNA without the RiboMinus rRNA removal step, typically, 20 to 60 µg of cRNA can be generated after cRNA cleanup.

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Why do you choose to use an rRNA-reduction strategy but not a poly-A mRNA-specific selection protocol?

It has been shown that a portion of the transcripts in total RNA do not necessarily contain Poly-A tails; therefore, they will be excluded by a poly-A RNA positive selection technique. The rRNA reduction approach will also make the protocol more robust in handling smaller amounts of total RNA samples as little as 1 µg.

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Why is Betaine added to the RiboMinus Hybridization Buffer?

Betaine increases the hybridization stringency. It equalizes the GC Tm and AT Tm, so the background, non-specific, non-rRNA hybridization to the RiboMinus probes due to high GC content may be reduced.

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What is the basic principle of the ribosomal RNA reduction procedure using the RiboMinus Human/Mouse Transcriptome Isolation Kit?

Four biotinylated LNA RiboMinus probes are designed to specifically bind to the abundant 18S and 28S rRNA species (2 probes each for 18S and 28S rRNA). Following hybridization of the biotinylated probes to the rRNA molecules in the total RNA sample, the rRNA is efficiently removed from the sample by the addition of the RiboMinus Magnetic Beads that are coated with streptavidin. The unbound fraction represents the RNA with rRNA species reduced. The sample is then concentrated before target labeling using the IVT cRNA Cleanup Kit. Consult the handbook included in the RiboMinus Kit (https://tools.thermofisher.com/content/sfs/manuals/ribominus_human_mouse_man.pdf) for more details.

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