Description
Hard-to-break samples including fatty and tough fibrous tissues (e.g., tendons or ligaments) pose significant challenges in the first step of DNA isolation. Conventional ways of sample rupture or enzyme digestion may not completely lyse those challenging samples. The SPINeasy® DNA Kit for Tissue (without Lysing Matrix) is a high-performance genomic DNA (gDNA) extraction kit which is based on silica-membrane spin-column technology. This kit enables convenient isolation of gDNA from a variety of solid tissues. The DNA purification protocol does not require any sophisticated equipment. Our specially formulated Buffer TD1 and Proteinase K provide effective lysis of various types of samples including tough, fibrous, and fatty tissues. The RNase A treatment (after lysis step) ensures the removal of potential RNA contamination. Furthermore, the subsequent addition of Buffer TD2 enables selective binding of DNA to Column S. The genomic DNA extracted from multiple types of tissues using this kit showed high integrity and purity, which can immediately be used for downstream applications, including long fragment PCR, real-time PCR, restriction enzyme digestion and sequencing.
Performance
Figure 1: Comparison of quality and quantity of gDNA extracted from various sample types using SPINeasy® DNA Kit for Tissue and Competitor Q. The gDNA yield, A260/A230 and A260/A280 ratio (from 15 mg of respective tissues) were measured with a spectrophotometer. The virtual gel image and DNA Integrity Number (DIN, in green, yellow, or red) were analyzed using Agilent Tapestation 4150.
Figure 2: Amplifiability of DNA extracted with SPINeasy® DNA Kit for Tissue (Without Lysing Matrix)
Upper panel: Agarose gel electrophoresis of PCR products amplified from gDNA isolated from various rabbit/mouse tissues using SPINeasy® DNA Kit for Tissue (Without Lysing Matrix). One microliter of extracted gDNA was amplified using rabbit or mouse-specific α-actin primers (1kb product). Reactions without DNA served as negative controls (NC).
Lower panel: Comparison of threshold cycles (Ct) of qPCR when amplification was performed using equal quantity of chicken liver, beef, and fish gill gDNA (25 ng) extracted with SPINeasy® DNA Kit and Competitor Q. Targets were amplified with SYBR green.