tRNA Microarray

Differential tRNA expression profoundly affects the entire dynamics of cellular translation, which favors or represses the expression of specific proteins. Due to their abundance of post-transcriptional modifications and their stable secondary structures, it is a challenge to measure the RNA levels directly or accurately using standard high-throughput sequencing or traditional RT-PCR techniques. Therefore, tailored and ingenious strategies must be devised to specifically assess cellular tRNA content. In recent years, the development of microarray technology has promoted the research of tRNA. In contrast to traditional bioassays, microarrays analyze predefined transcripts/genes by hybridization, allowing simultaneous measurement of tens of thousands of tRNAs. tRNA microarray is a novel, straightforward, and cost-effective method for the rapid and accurate measurement of tRNA abundance in biological samples, and has become an important part of biological and biomedical research. BOC Sciences provides tRNA Microarray services to help you understand important biochemistry parameters of translational in vivo by accurately measuring the absolute amounts of tRNA.

Fig 1. Microarray images of tRNA. (Dittmar KA, 2006)

tRNA Microarray Services

BOC Sciences' tRNA microarray consists of 40 full-length tDNA probes that recognize all 54 nuclear-encoded human isoreceptor tRNAs. Deacetylated tRNAs are selectively labeled with a fluorescent oligonucleotide hairpin, and differently labeled tRNAs from different conditions or cell/tissue types are labeled with the oligonucleotide hairpins with two different colors. The tRNAs are then mixed and hybridized onto the tRNA microarray. The ratio between the two different fluorescent signals serves as a relative comparison of tRNA between the two samples.

Fig 2. Schematics of fluorescence labeling of tRNA in a total RNA mixture. (Dittmar KA, 2006)

Features of Our tRNA Microarray

BOC Sciences provides microarray analysis of tRNA samples labeled in vitro with fluorochromes or radioisotopes, which are applicable to most eukaryotic or prokaryotic organisms.

• Simultaneous profiling of the tRNA

Our tRNA microarray uses direct end-labeling and intelligent probe design to realize simultaneous detection and quantification of tRNA on one chip in one experiment, providing important expression information and biomarker applications for the study of tRNA regulation.

• High sensitivity, specificity and accuracy

Our employment of direct end-labeling and hybridization with high-affinity probes enables very high sensitivity, specificity, and accuracy even for tRNAs at low-abundance. We can reliably distinguish tRNA isoacceptors with a sequence difference of more than 8 nucleotides. Metrics such as reproducibility, sensitivity, and specificity are thoroughly evaluated, and are found to meet or exceed current standards for tRNA analysis.

• Straightforward and simplified procedure

Our tRNA microarray service overcomes biases from RNA modification, RNA folding hindrance, reverse transcription block, PCR amplification and inaccurate small RNA-seq analysis.

• Low tRNA sample amount requirements

Because the direct end-labeling chemistry is less susceptible to RNA loss which is caused by the RNA pretreatment process, our microarray service significantly reduces the need for large amounts of modified tRNA, requiring as little as 100 ng of tRNA sample. The low sample size requirement presents an opportunity for research projects where the samples are rare or in limited supply.

• Tolerant to lower quality tRNA samples

The direct RNA end-labeling in BOC Sciences' tRNA microarray analysis is relatively insensitive to nucleotide damage in the substrate RNA sequence because it does not rely on cDNA replication by reverse transcription. Therefore, our tRNA microarray is particularly advantageous for preserved or chemically treated samples or degraded samples such as serum/plasma/biological fluid RNA, FFPE RNA.

Application of tRNA Microarray

• Measure the absolute amounts of tRNA and tRNA charging levels.
• Identify low-abundance species such as tRNAs associated with polysomes.
• Study the biological relevance of tRNA expression changes, such as physiological and pathological processes.