Description : The EZgeneTM Plant gDNA maxi Kit is designed for efficient recovery of genomic DNA up to 60 kb in size from fresh and dried plant tissue samples. Up to 2 g of wet tissue (or 500 mg dry tissue) can be processed in one column. The system combines the reversible nucleic acid-binding properties of the matrix with the speed and versatility of spin column technology to eliminate polysaccharides, phenolic compounds, and enzyme inhibitors from plant tissue lysates. Purified DNA is suitable for PCR, restriction digestion, and hybridization techniques. There are no organic extractions thus reducing plastic waste and hands-on time to allow multiple samples to be processed in parallel. If using the EZgeneTM Plant gDNA Kit for the first time, please read this booklet to become familiar with the procedures. Samples are homogenized and lysed in a high salt buffer. The DNA is bound to the column while proteins and other impurities are removed by wash buffer. The purified DNA is suitable for downstream applications such as endonuclease digestion, thermal cycle amplification, and hybridization techniques. All components of the EZgeneTM Plant gDNA maxi Kit are stable for at least 12 months from date of purchase when stored at 22 oC -25 oC. RNase A should be stored at 4 oC. During shipment or storage in cool ambient conditions, precipitates may form in Buffer P1 and Buffer P2. It is possible to dissolve such deposits by warming the solution at 37 oC, though we have found that they do not interfere with overall performance. Product GD2614-00 GD2614-01 GD2614-02 DNA Maxi Columns 2 10 25 50 mL collection tubes 2 10 25 Buffer P1 40 mL 180 mL 500 mL Buffer P2 10 mL 40 mL 100 mL Buffer P3 10 mL 40 mL 100 mL RNase A 110 μL 550 μL 1.3 mL Buffer KB 25 mL 110 mL 280 mL DNA Wash Buffer 30 mL 90 mL 2x 90 mL Elution Buffer 10 mL 50 mL 125 mL User Menu 1 1 1 Prepare all components and get all necessary materials ready by examining this instruction booklet and become familiar with each steps. GD2614-00 Add 70 mL absolute (96%-100%) ethanol GD2614-01 Add 210 mL absolute (96%-100%) ethanol GD2614-02 Add 2 x 210 mL absolute (96%-100%) ethanol per bottle A. Dry Specimens (Page 4) For processing ≦500 mg powdered tissue. B. Fresh/Frozen Specimens (Page 6) For processing ≦2 g fresh (or frozen) tissue. Possible reasons Suggestions Clogged column Carry-over of debris. Make sure no particulate material is transferred. Sample too viscous. Do not exceed suggested amount of starting material. Alternatively, increase amounts of Buffers P1 and P2 and use two or more columns per sample. Low DNA yield Incomplete disruption of starting material. For both dry and fresh samples, obtain a fine homogeneous powder before adding Buffer P1. Poor lysis of sample. Decrease amount of starting material or increase amount of Buffers P1, P2 and P3 DNA remains bound to column. Increase elution volume to 3 mL and incubate on column at 65℃ for 5 min before centrifugation. DNA washed off. Dilute Wash Buffer by adding appropriate volume of absolute ethanol prior to use ( page3 ). Problems in downstream applications Salt carry-over. DNA Wash Buffer must be at room temperature. Ethanol carry-over Following the second wash spin, ensure that the column is dried by centrifuging 2 min at maximum speed.Introduction
Overview
Storage and Stability
Kit Contents
Before Starting
Important
Trouble Shooting Guides
Problems
- Model: GD2614-00
- Manufactured by: Biomiga