Description :

Introduction

The EZgene^TM 96-Well Blood DNA Kit allows rapid and reliable isolation of high-quality genomic DNA /viral DNA in a high-through-put 96-well format from a wide variety of samples including fresh, frozen, or anticoagulated whole blood, serum, plasma, bone marrow, body fluids, lymphocytes and cultured cells. Blood DNA is bound to Biomiga's ezBind matrix while proteins and other unwanted impurities are removed by two rapid wash. Pure DNA is then eluted from the matrix with Elution Buffer or ddH₂O. Purified DNA is suitable for PCR, restriction digestion, and hybridization techniques. The DNA binding capacity per well is 50 µg.

Storage and Stability

All components of the EZgene^TM 96-Well DNA Kit are stable for at least 12 months from date of purchase when stored at 22-25°C. Buffer BL may form precipitates in cool ambient conditions, Warm up the bottle at 37°C to dissolve before use. Store Protease K at -20°C.

Safety Inforamtion

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

Materials to be provided by user

•  Laboratory centrifuge capable of at least 5,000 x g equipped with swinging bucket rotor.
•  Rotor adapter for deep well microplates
•  Water bath equilibrated to 65 °C
•  Absolute (96%-100%) ethanol
•  Multichannel pipet with tips
•  Optional: RNase A stock solution (20 mg/mL)
•  Incubator or vacuum oven preset at 65°C

Before Starting

Please read the entire manual to become familiar with the EZgene^TM 96-Well Blood DNA kit procedure.

1. Prepare a Protease K stock solution with Elution Buffer and aliquot into adequate portions. Store each aliquot at -20 °C and thaw before use. Each sample will require 25 μL of this solution.

• l GD2815-00 Dissolve with 2.5 mL Elution Buffer
• l GD2815-01 Dissolve with 10.0 mL Elution Buffer
• l GD2815-02 Dissolve with 50.0 mL Elution Buffer

2. Dilute DNA Wash Buffer Concentrate with absolute ethanol as follows and store at room temperature.

• l GD2815-00 Add 160 mL (96%-100%) ethanol
• l GD2815-01 Add 640 mL (96%-100%) ethanol
• l GD2815-02 Add 640 mL (96%-100%) ethanol per bottle

3. Preheat Elution Buffer at 65 °C.

4.Adjust the volume of samples to 250 μL. For samples smaller than 250 μL, add appropriate volume of PBS to 250 μL. For samples larger than 250 μL, split each sample into two 250 μL aliquots and use two wells of the 1.2 mL round well plate for lysis. Load the combined lysates into each well of the 96-Well DNA Plate.

Storage of Blood Samples

Storage of blood samples without previous treatment leads to reduced yields of genomic DNA. For the best result, blood samples should be treated as follows,

• l For short-term storage (up to a week), collect blood in tubes containing EDTA as anticoagulant, and store at 4 °C.
• l For long-term storage, collect blood in tubes contain an anticoagulant and store at -70°C. Thawed frozen blood sample at 37°C with gently agitation before use.

Determination of Yield and Quality

The total DNA yield can be calculated as:

[DNA] = (Absorbance260) x (0.05 μg/μL) x (Dilution factor)

The quality of DNA can be assessed by measuring absorbance at A_260/280. A ratio of (A260/A280) of 1.8-1.9 corresponds to 90%-95% purity.