Description :

Introduction

The EZgene^TM Mollusc gDNA Kit is designed to extract genomic DNA up to 60 kb in size from molluscs, insects, arthropods, roundworms, flatworms, and other invertebrate tissue samples rich in mucopolysaccharides. The method is suitable for invertebrates frozen or preserved in alcohol or DNE solution, and good results can be obtained with formalin preserved material.

Samples are homogenized and lysed in a high salt buffer and extracted with chloroform to remove mucopolysaccharides. Following a rapid alcohol precipitation step, binding conditions are adjusted and DNA further purified using ezBind DNA spin columns. While proteins and other contaminants are removed by wash buffer, high quality genomic DNA is eluted with elution buffer or sterile water. The purified genomic DNA is suitable for downstream applications such as Southern Blot, restriction digestion, and PCR.

Storage and Stability

All components of the EZgene^TM Mollusc gDNA Kit, except the Proteinase K and RNase A should be stored at 22^oC -25^oC. Once reconstituted in water, Proteinase K should be stored -20^oC.Under these conditions, DNA has successfully been purified and used for PCR after 24 months of storage. During shipment, or storage in cool ambient conditions, precipitates may form in some buffers. It is possible to dissolve such deposits by incubation the solution at 65 ^oC. Store RNase A at -20 ^oC. All EZgene^TM Mollusc gDNA Kit components are guaranteed for at least 12 months from the date of purchase when stored at 22 ^oC -25 ^oC.

Binding Capacity

Each ezBind DNA column can bind approximately 100 μg DNA. Use less than 30 mg of sample per column.

Kit content

Before Starting

Please read the entire booklet to become familiar with the EZgene protocol.

Dilute DNA Wash Buffer with 100% ethanol as follows:

Reconstitute Proteinase K stock solution. Vortex vial briefly prior to use. We recommend that you aliquot and store vials of reconstituted protease at -20 ℃.

Determination of DNA Quality and Quantity

Dilute a portion of the eluted material approximately 10-20 fold in ddH₂O Measure absorbance at 280 nm and at 260 nm to determine the A260/A280 ratio. Values of 1.7-1.9 generally indicate 85%-90% purity. The concentration of DNA eluted can be determined as follows:

Concentration = 50 μg/mL x Absorbance260 x {Dilution Factor}

Operating Protocol