Introduction
The Blood DNA Kit provides a rapid and easy method for the isolation of genomic DNA from 1 µl-1 ml fresh, frozen, and anti-coagulated whole blood. The method can also be used for preparation of genomic DNA from buffy coat, serum, and plasma. The kit allows single or multiple, simultaneous processing of samples in less than 90 minutes.
The use of mini-columns facilitate the binding, washing, and elution steps thus enabling multiple samples to be simultaneously processed. For more information on columns, click Flexible Multi-Format Columns.
Principle
Blood DNA Kits use the reversible nucleic acid-binding properties of HiBind® matrix, combined with the speed of mini-column spin technology. A specially formulated buffer system allows genomic DNA up to 60 kb to bind to the matrix. Samples are first lysed under denaturing conditions and then applied to the HiBind® spin columns to which DNA tightly binds, enabling cellular debris, hemoglobin subsequently eluted in sterile deionized water or low salt buffer.
Procedure
OB Protease or proteinase K and RNase are added to cells to efficiently degrade proteins and RNA present in the blood. After an incubation period, isopropanol is added to the sample to precipitate DNA from solution to enable binding to the HiBind® spin column. After addition of isopropanol solution is applied to column and centrifuged to bind DNA to column. Once bound to the matrix in the column DNA is subjected to multiple wash steps and eventually eluted using sterile water of elution buffer.
Blood DNA Kit Protocol
Applications
The DNA isolated using the Whole Blood Genomic DNA Isolation Kit is free of all contaminants such as RNA, protein, and metabolites, and has an 260/280 absorbance ratio between 1.7 and 1.9, making it well suited for use in the following applications:
- Restriction Digestion
- Southern blotting
- PCR amplification
