Clariom™ D Assay，人类

Please refer to the Microarray Reagent Guide for Arrays and Expression Kits to match the correct reagents your array.

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Tough-Spots® labels are small adhesive stickers used to temporarily seal the backs of cartridge arrays during the overnight hybridization step. They are required to prevent loss of volume due to evaporation through the septa. We recommend using Tough-Spots® labels on Rolls from USA Scientific (Item No. 9185-0000)

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Proper hybridization volume is critical to obtaining an even signal across a given array. Too little volume can lead to black circles in the middle of the array. Too much volume can leak out of the back of the array. The correct hybridization volume leaves enough room for a small air bubble to circulate around the array surface during the overnight hybridization. Here are the recommended hybridization and fill volumes based on the array format:
Array Format; Hybridization Volume; Fill Volume
- 49 Format (Standard); 200 µL; 250 µL
- 64 Format; 200 µL; 250 µL
- 100 Format (Midi); 130 µL; 160 µL
- 169 Format (Mini); 80 µL; 100 µL
- 400 Format (Micro); 80 µL; 100µL

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Clariom D arrays have probes that cover all known regions of transcription including probes in overlapping regions from both strands. To obtain strand-specific information from the Clariom D arrays, the WT Pico and WT Plus reagents (which are strand-specific) must be used. This is important because without strand-specific reagent, it would not be possible to decipher the source strand of DNA, which makes it challenging to untangle true gene- and exon- level expression and alternative splicing events.

Strand-specificity is significantly less important for customers interested in gene-level only information (i.e., those using Clariom S) as compared to customers who want to understand the complexities of the whole transcriptome including identifying antisense transcripts and ncRNA (i.e., those using Clariom D). While strand-specificity is less important for gene-level expression only, probes within regions of overlapping transcription from both strands are avoided in the Clariom S array design (unlike Clariom D). This is important because if Clariom S did not preserve strand-specificity, there could be an overestimation of gene-level expression causing false positive or negative results. With Clariom S having a "stranded" design, it does not necessarily need a strand-specific reagent kit.

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TAC 4.0 includes two algorithms for identifying alternative splicing events: the TAC 2.0 algorithm and the new EventPointer. Algorithmic determination of alternate splicing remains a challenging problem. TAC 4.0 supports two different approaches that have different sets of strengths and weaknesses. After considerable testing, the new TAC 4.0 “'Event Score” leverages both previous TAC 2.0 event estimation score and Event Pointer p-value and sorts the most likely alternative splicing events to the top. Of course, the TAC 2.0 event score and EventPointer p-values remain individually available.

Find additional tips, troubleshooting help, and resources within our Microarray Analysis Support Center.