복잡한 동물 조직에서 총 RNA 추출의 기술적 최적화: 마그네틱 비드 방법을 기반으로 한 미니 추출 프로토콜의 평가

1.Abstract

Animal visceral tissues (such as liver, kidney, 마음, and intestinal tissues) are rich in endogenous nucleases, 다당류, 지질, and connective tissue components. As a result, the extraction of total RNA from these tissues has long faced technical challenges such as a high risk of degradation, low product purity, and unstable yields. The traditional phenol-chloroform extraction method is not only cumbersome and time-consuming but also involves toxic organic reagents, making it difficult to meet the demands of high-throughput and automated experiments.

Based on application data from the Shanghai Lingjun Biotech 범용 총 RNA 추출 키트 (마그네틱 비드 방식) in various animal visceral samples, this paper systematically evaluates its performance in small-volume extractions. The results indicate that the kit effectively removes protein and polysaccharide contamination. The A260/A280 and A260/A230 ratios of the extracted RNA both meet the quality requirements for downstream molecular biology experiments, and the yield is sufficient for various applications, such as RT-qPCR and transcriptomic sequencing, providing a safe and efficient solution for RNA extraction from complex tissue samples.

2.소개

The integrity and purity of ribonucleic acid (RNA) are critical prerequisites for molecular biology experiments such as gene expression analysis, transcriptomic sequencing, and non-coding RNA research. Animal visceral tissues—particularly the liver, kidneys, 마음, and intestines—are typically considered “difficult samples” for RNA extraction. The liver is rich in glycogen and endogenous RNases; kidney and intestinal tissues contain large amounts of polysaccharides and proteolytic enzymes; and the heart, due to the dense structure of its muscle fibers, does not lysis sufficiently. These factors can all lead to RNA degradation, low yield, or the co-purification of impurities, which in turn affect reverse transcription efficiency and the reliability of quantification results.

최근에는, the magnetic bead method for nucleic acid extraction has gradually emerged as an alternative to traditional centrifugation column and organic solvent methods, thanks to its advantages of operational safety, ease of automation, and consistent product quality. This study evaluated the performance of a universal total RNA extraction kit based on the magnetic bead method, developed by Shanghai Lingjun Biotechnology, in visceral samples from pigs and mice, providing a systematic technical reference for RNA extraction from such samples.

3.Product Technical Principles

This kit is based on surface-modified magnetic bead technology and utilizes the selective adsorption of nucleic acids by magnetic beads in a specific buffer system to purify RNA. The extraction process includes: releasing total RNA from the sample using a lysis buffer; specific binding of nucleic acids to the magnetic beads; removal of protein and salt ion impurities using wash buffers; and elution with a nuclease-free wash to obtain high-purity RNA. This system does not require the use of organic solvents such as phenol and chloroform, thereby avoiding the health risks and waste disposal burdens associated with traditional methods. 추가적으로, the magnetic bead method prevents column clogging caused by sample particulates during centrifugation-based filtration, and offers flexible elution volumes (30–100 μL), making it suitable for the concentration and recovery of low-concentration RNA samples.

4.Experimental Data and Analysis

Approximately 5–10 mg each of fish liver, 장 조직, gills, and heart were collected and extracted using the Lingjun Biotech Universal Total RNA Small-Volume Extraction Kit. The results are shown in the table below.

아니요.	견본	260/280	260/230	농도. (ng/μL)
1	5 mg 생선 간	1.92	1.54	272.95
2	10mg Fish guts	1.84	2.04	119.15
3	10mg Fish gill	1.92	2.13	276.15
4	10mg 물고기 심장	1.89	1.89	113.70

Electrophoresis Results：

보시다시피, the A260/280 ratios are above 1.9, the concentrations are all above 100 ng/μL, 메인 밴드가 선명하게 보입니다..

5.Discussion and Recommendations for Application

5.1 Technical Advantages of the Magnetic Bead Method

Compared to the traditional Trizol method, the magnetic bead method significantly reduces processing time (the entire process can be completed in under 40 분) and minimizes the risk of RNA degradation due to exposure during the procedure. 게다가, the magnetic bead method is compatible with automated nucleic acid extraction platforms, 처리량이 많은 시료 처리에 적합합니다..

5.2Extraction Strategy

Small-volume extraction is suitable for routine analyses such as RT-qPCR, cDNA library construction, and Northern blot; it requires only a small amount of sample and is ideal for the initial screening of precious samples. For tissues with high fat content or high RNase levels (such as the pancreas), the amount of lysis buffer can be appropriately increased or an RNase inhibitor can be added in advance to optimize extraction efficiency.

6.결론

The Shanghai Lingjun Biotech Universal Total RNA Extraction Kit (마그네틱 비드 방식) demonstrates excellent purity, 생산하다, and ease of use in the extraction of RNA from animal visceral tissues. This system provides a technical solution that combines efficiency and quality for RNA research on complex biological samples, and the resulting RNA is suitable for a wide range of downstream applications, from RT-qPCR to transcriptomic sequencing.

공급자

상하이 링쥔 생명공학 유한회사, 주식회사에 설립되었습니다 2016 생체자성재료 및 핵산추출시약 전문제조업체입니다..

핵산 추출 및 정제 분야에서 풍부한 경험을 보유하고 있습니다., 단백질 정제, 세포 분리, 화학발광, 및 기타 기술 분야.

우리의 제품은 많은 분야에서 널리 사용됩니다., 의료 테스트와 같은, 유전자 검사, 대학 연구, 유전자 육종, 등. 우리는 제품을 제공할 뿐만 아니라 OEM도 수행할 수 있습니다., ODM, 그리고 다른 필요. 관련된 필요사항이 있는 경우, 저희에게 연락하게 자유롭게 느끼십시오 .