Description :

Introduction

The EZgene^TM Blood gDNA kit provides a fast and easy method for isolating gDNA from blood. The system utilizes the reversible nucleic acid-binding properties of Biomiga's ezBind membrane and the speed of spin column technology to yield high quality gDNA with the OD_260/280 ratio of 1.8-2.0. Up to 250 μL of fresh, frozen or anticoagulated whole blood can be readily processed at one time. This DNA Kit can also be used for the preparation of genomic DNA from buffy coat, serum, plasma, saliva, buccal swab and other body fluids. Purified DNA is ready for applications such as PCR, Southern Blotting, and Restriction Digestion.

The binding capacity per column is 100 μg of gDNA. Use less than 250 μL of whole Blood or buffy coat is recommended.

Storage and Stability

All EZgene^TM Blood gDNA Kit components are guaranteed for at least 12 months from the date of purchase when stored as follows:

• Reconstituted Protease K and store at -20°C.
• All other materials at room temperature (22-25°C).

Safety Information

Buffer BL contains chaotropic salts, which may form reactive compounds when combines with bleach, Do not add bleach or acidic solutions directly to the preparation waste, ware gloves and protective eyewear when handling.

Kit Contents

Before Starting

The kit is designed to be simple, fast, and reliable provided that all steps are followed diligently. Please read the entire booklet and get all necessary supplies and equipments.

Important

• Reconstitute Protease K with 110 μL (GD2311-00), 1.3 mL (GD2311-01), or 5 x 1.3mL (GD2311-02) Elution Buffer and vortex the vial briefly before use. Aliquot and store vials of reconstituted Protease K at -20°C.
• Dilute DNA Wash Buffer with absolute ethanol as follows:

 Add 8 mL (GD2311-00) or 60 mL (GD2311-01) or 96 mL (GD2311-02) of absolute ethanol to each bottle. The final concentration is 80%.

• Under cool ambient conditions, precipitates may form in Buffer BL. In case of such an event, heat the bottle at 37°C to dissolve before use.

Materials provided by user

• Tabletop microcentrifuge
• Sterile 1.5 mL centrifuge tubes
• Water bath
• Absolute ethanol
• PBS Buffer

Determination of Yield and Quality

The total DNA yield can be determined by a spectrophotometer using deionized water, Tris-HCl Buffer, or Elution Buffer as a blank. Dilute the DNA in TE buffer and calculate the concentration using the following equation:

[DNA] = (Absorbance ₂₆₀) x (0.05 μg/μL) x (Dilution Factor)

The quality of DNA can be assessed by measuring absorbance at both 260 nm and at 280 nm. A _{260/ 280} ratio of 1.7-1.9 corresponds to 85%-95% purity. Expected yields will range from 4 μg - 12 μg of DNA per 250 μL of whole blood, depending on the source of the sample, its age, and the method of storage. Yields are generally 5-fold higher with buffy coat samples.

Operating Protocol