Mammalian Genomic DNA Miniprep Kit

For purification of genomic DNA from a variety of mammalian sources

Description: The GenElute Mammalian Genomic DNA Purification Kit provides a simple and convenient way to isolate pure, high molecular weight DNA from a variety of mammalian sources (Figure 1 and Figure 2). These kits use a silica-based membrane, specially selected for genomic DNA purification, in a convenient spin column format (G1N10, G1N70, G1N350).

Procedure

Mammalian cells and tissues are lysed with a chaotropic salt-containing buffer to ensure denaturation of macromolecules. DNA is bound to the spin column membrane and the remaining lysate is removed by centrifugation. A filtration column is used to remove cell debris. After washing to remove contaminants, the DNA is eluted with buffer into a collection tube. The purified DNA may be used in many applications such as sequencing, cloning, blotting, restriction digestion, ligation, and PCR.

Features and Benefits

• Typical DNA yields of 25 µg from 2 x 106 cultured cells or 30 µg from 25 mg of tissue (Figure 4)
• Preparation time is only 20 minutes after lysis
• Purified genomic DNA has A260/A280 ratios between 1.6 and 1.9
• No need for mechanical homogenization
• 40% more purification preps offered than other leading supplier

Storage

Room Temperature
R: 20/21/22 S: 26-36

Application

Purified DNA may be used in many applications such as sequencing, cloning, blotting, restriction digestion (Figure 3), ligation, and PCR (Figure 4).

Table 1. Yields produced using GenElute Mammalian Genomic DNA Purification Kits (with RNase and Proteinase K treatment).

Material	Amount	Typical Yield
Jurkat Cells (human)	2 x 106 Cells	5 - 10 µg
HEK293 Cells (human)	2 x 106 Cells	10 - 20 µg
HeLa Cells (human)	2 x 106 Cells	15 - 25 µg
Mouse Pancreas Tissue	20 mg	10 - 25 µg
Mouse Spleen Tissue	10 mg	10 - 25 µg
Mouse Thymus Tissue	16 mg	10 - 25 µg
Mouse Lung Tissue	20 mg	5 - 15 µg
Mouse Brain Tissue	16 mg	5 - 15 µg
Mouse Kidney Tissue	20 mg	10 - 25 µg
Mouse Liver Tissue	25 mg	10 - 30 µg

Genomic DNA Purified from
Mammalian Tissues

Figure 1. Genomic DNA purified from mammalian tissues. Purified genomic DNA, from the indicated mouse tissues, was isolated with the GenElute™ Mammalian Genomic DNA Miniprep Kit. Genomic DNA (200 ng/lane) was analyzed on a 0.8% agarose gel to illustrate yield and integrity. Markers are lambda DNA digested with Hind II. (Product No. D9780)

Genomic DNA Purified from Cells using GenElute Mammalian Genomic DNA Purification Kit

Figure 2. Genomic DNA purified from mammalian cells. Purified genomic DNA was isolated with the GenElute Mammalian Genomic DNA Purification Kit from 2 x 106 cells of the sources indicated. The genomic DNA (200 ng/lane) was analyzed on a 0.8% agarose gel. Marker is a Lambda Hind III digest (0.5-23 kb).

Genomic DNA Purified with GenElute Mammalian Genomic DNA Purification Kit is Suitable for Restriction Enzyme Digestions.

Figure 3. Genomic DNA (200 ng) was digested with EcoR I (10 units in 20 µL at 37°C for 4 hours) followed by electrophoresis (150 ng/lane) on a 0.8% agarose gel. The undigested DNA was incubated under the same conditions, but without EcoR I. Marker is a Lambda Hind III digest (0.5-23 kb).

Yield Comparison

Figure 4. Yields of genomic DNA compared with leading Supplier Q using 2 x 200 µL elutions.