Among the parameter transcriptomes, RPKM>1 is considered to be gene expression. This threshold is recommended by mainstream magazines and can also well reflect gene expression levels. 

Cluster analysis is used to judge the expression patterns of differential genes under different experimental conditions; By grouping genes with the same or similar expression patterns into clusters, the functions of unknown genes or unknown functions of known genes can be identified; Because these similar genes may have similar functions or participate in the same metabolic process or cellular pathway. 

There are many methods to verify Small RNA in subsequent experiments: 

1.  qPCR is used to verify the expression of miRNA; 

2. For RNA with methylation modification enriched by RIP technology or RNA specifically binded to the target protein, the former shall be carried out the RIP-seq, and compared and analyzed with the genome in order to obtain position information specifically binding to the target protein on miRNA, etc.; 

3. Dual-Luciferase reporter assay system  MiRNA mainly acts 3'UTR on the target gene, and the 3 'UTR region of the target gene can be constructed behind luciferase, the reporter gene, in the carrier. The inhibiting effect of miRNA on the target gene can be quantitatively reflected by the changes on expression of the reporter gene (monitoring the changes in luciferase activity), after comparing the overexpression or interference with miRNA, and, this is, we can judge whether miRNA binds to the target gene via the fluorescence value. 

Small Small RNAの後続の実験検証の方法はたくさんあります。

1.qPCR：miRNAの発現を検証します

2.qPCR：RIP技術の濃縮によりメチル化修飾付きRNAまたは目的タンパク質と特異的に結合したRNAを得、濃縮により得られたRNAに対してハイスループット配列決定を行い、ゲノムと比較分析を行い、miRNAにおける目的タンパク質と特異的に結合した位置の情報などを得ます。

3.Dual-Luciferase報告システム

   miRNAは主に標的遺伝子に作用する3′UTRが作用することにより、目的遺伝子3′UTR領域をベクター中の報告遺伝子luciferaseの下流に構築し、過発現あるいは妨害miRNAを比較することにより、遺伝子発現の変化(ルシフェラーゼの活性変化のモニタリング)を報告し、目的遺伝子に対するmiRNAの抑制作用を定量的に反映し、したがって、miRNAが標的遺伝子と結合するかどうかは蛍光値で判断できます。

Blood RNA stabilizer (DP440) can be used for storage of whole blood samples. Mixing ratio of whole blood: stabilizer=1:3, it can be stored for 3 months at -20℃. It can be stored for 3 ~ 5 days at normal temperature. Room temperature transportation can be selected. 

Many genome regions have positive and negative strands of transcripts. Antisense transcription is a feature of eukaryotic genes and an important regulation mode. To more

accurately obtain the structure of the gene and gene expression information, the use of ssRNA-Seq (Strand-Specific) can retain the directional information of RNA, and can determine whether the transcript comes from positive or negative strand.