Application

DNA Isolation Kit for Cells and Tissues has been used to isolate DNA from a wide variety of starting materials. The isolated DNA is suitable for many molecular biology applications:Genomic Southern blottingSequencingRestriction digestionPCR/long PCRCloning

Components

Cellular Lysis Buffer Proteinase K Solution RNase Solution Protein Precipitation Solution

Features and Benefits

Obtain increased yields of DNA in less than 2.5 hours.Benefit from a simple, straightforward procedure (compared with column based methods).Increase safety and convenience.Eliminate the use of chaotropic salts, anion-exchange columns, and hazardous organic solvents.Reduce purification time.All required reagents are included in the kit.

General description

The DNA Isolation Kit for Cells and Tissues provides medium- and large-scale preparation of purified genomic DNA ranging in size from 50 to 150 kb. Remove contaminating RNA and proteins from a wide variety of biological specimens (mammalian tissue, cultured cells, yeast, gram-negative bacteria or mouse tail).

Other Notes

For life science research only. Not for use in diagnostic procedures.

Purity of isolated DNA: Average A ₂₆₀ / A ₂₈₀ ratio: 1.7 - 1.9Isolation of High Molecular Weight DNAThe kit simplifies the isolation of 50 to 150 kb genomic DNA.

Figure 1: Amplification of both short and long DNA fragments from genomic DNA prepared with the DNA Isolation Kit for Cells and Tissues. Genomic DNA was isolated from a variety of sources and then amplified with either Taq DNA Polymerase, the Expand High Fidelity PCR System, or the Expand Long Template PCR System.Lanes 2, 3: Human DMD fragment (268 bp) and mouse c-myc fragment (580 bp), amplified with Taq DNA PolymeraseLanes 4, 5, 7, 8: Human c-myc fragment (1.2 kb), mouse β2-microglobulin fragment (3.6 kb), bovine lysozyme gene fragment (6.9 kb), and human tPA gene fragment (9.3 kb), all amplified with the Expand High Fidelity PCR SystemLanes 6, 9, 10: Mouse α-2 collagen gene fragments (5.6 kb and 10.4 kb) and human β-globin fragment (23 kb), amplified with the Expand Long Template PCR SystemLanes 1, 11 : DNA Molecular Weight Markers VI and II

Preparation Note

Sample material is homogenized in Cellular Lysis Buffer in the presence of a strong anionic detergent and proteinase K. RNA is eliminated by RNase treatment and proteins are removed by selective precipitation and centrifugation. The purified DNA is finally recovered from solution by isopropanol precipitation.

Quality

Each lot of the DNA Isolation Kit for Cells and Tissues is tested for the absence of DNase contamination. Function tests to purify DNA from bovine liver, followed by specific amplification with the Expand High Fidelity PCR System were performed.