Genomic DNA Isolation Kit

Supporting Data

Figure 1. Isolation of High Quality Genomic DNA. Genomic DNA was isolated from various sample types using Norgen's Genomic DNA Isolation Kit. Lanes 1-3 contain genomic DNA that was isolated from four different samples containing 5 x 10⁵ HeLa cells, while lanes 4 and 5 contain genomic DNA that was isolated from 5 mg of heart tissue. All the purified genomic is of the highest quality and integrity. Lane M is the Norgen HighRanger 1kb DNA Ladder. For all purified DNA, 10 µL of each 200 µL elution were resolved on a 1X TAE, 1% agarose DNA gel.

Figure 2. Full Compatibility with Digests. Genomic DNA was isolated from 10 mg of kidney tissue using Norgen's Genomic DNA Isolation Kit, and subsequently digested. Twenty microliters of the 200 µL eluted DNA were used in a 50 µL restriction digestion with 10 units of either Hind III or Nde I (overnight at 37°C). Fifteen microliters of the digested DNA were then loaded on a 1X TAE, 1% agarose gel. Lane 1 contains undigested Norgen-isolated DNA, while Lanes 2 contain the genomic DNA digested with Hind III and Lanes 3 contain the genomic DNA digested with Nde I. Lane M is Norgen's HighRanger 1kb DNA Ladder.

Kit Specifications
Column Binding Capacity	25 µg
Average Yield:* HeLa Cells (1 x 10⁶) Tissue (from 10 mg kidney)	8 µg 10 µg
Maximum Amount of Starting Material: Animal Tissues Cultured Cells Bodily Fluids (blood, saliva) Viral Suspension	20 mg 3 x 10⁶ cells 150 µL 150 µL
Time to Complete 10 Purifications	80 minutes

* Yield will vary depending on the type of sample processed

Storage Conditions and Product Stability
The Proteinase K should be stored at -20°C upon arrival and after reconstitution. All other solutions should be kept tightly sealed and stored at room temperature. This kit is stable for 2 years after the date of shipment.

Component	Cat. 24700 (50 preps)	Cat. 24750 (100 preps)	Cat. 24770 (250 preps)
Digestion Buffer A	25 mL	2 x 25 mL	5 x 25 mL
Buffer SK	30 mL	2 x 30 mL	5 x 30 mL
Wash Solution A	18 mL	2 x 18 mL	5 x 18 mL
Elution Buffer B	30 mL	2 x 30 mL	5 x 30 mL
Proteinase K	12 mg	2 x 12 mg	5 x 12 mg
Spin Columns	50	100	250
Collection Tubes	50	100	250
Elution Tubes (1.7 mL)	50	100	250
Product Insert	1	1	1

Documentation

PROTOCOLS

(24700) Genomic DNA Isolation Kit - Protocol (50 prep)

(24750) Genomic DNA Isolation Kit - Protocol (100 prep)

(24770) Genomic DNA Isolation Kit - Protocol (250 prep)

SAFETY DATA SHEETS

24400 - Plant RNA_DNA Purification Kit - SDS

24750 - Genomic DNA Isolation Kit (100 Prep) - SDS

24770 - Genomic DNA Isolation Kit (250 Prep) - SDS

FAQs

Spin Column

I noticed the column was clogged. What causes this?

Column clogging may occur due to the sample being too large. Do not exceed the recommended amount of starting materials. The amount of starting material may need to be decreased if the column shows clogging below the recommended levels. Clogging can also be alleviated by increasing the g-force and/or centrifuging for a longer period of time until the lysate passes through the column.

Why is the lysate very gelatinous prior to loading onto the column?

Lysate can be more gelatinous prior to loading onto the column due to the following factors:

The lysate solution mixture is not homogeneous. To ensure a homogeneous solution, vortex for 10-15 seconds before applying the lysate to the spin column.
Maximum number of cells or amount of tissue exceeds kit specifications. Refer to specifications to determine if the amount of starting material falls within kit specifications.

Why do I have a low genomic DNA yield?

A low genomic DNA yield may be caused by:

Improper storage of samples. Tissue samples and cell pellets may be frozen and stored at -20°C or -70°C. Repeated freezing and thawing of stored samples should be avoided, as this may lead to decreased yields of DNA.
Incomplete lysis of cells. Extend the incubation time of Proteinase K digestion or reduce the amount of tissue or cells used for lysis.
The DNA elution is incomplete. Ensure that centrifugation at 14,000 x g is performed after the 3,000 x g centrifugation cycle, to ensure that all the DNA is eluted.

Why is the genomic DNA sheared?

Shearing of genomic DNA can be due to the following reasons:

The genomic DNA was handled improperly. Pipetting steps should be handled as gently as possible. Reduce vortexing times during mixing steps (no more than 10-15 seconds).
Improper storage of sample. Repeated freezing and thawing of stored samples should be avoided as this may lead to decreased DNA size.
The sample is old. Sheared DNA may be obtained from old tissue or cell samples. Fresh samples are recommended for maximum genomic DNA yield.

Can this kit also isolate mitochondrial DNA (mtDNA)?

Yes, mitochondrial DNA (mtDNA) can be successfully isolated using this kit. Please contact our Tech support team at support@norgenbiotek.com and ask for reference publications.