Description : The EZgeneTM Plant gDNA Kit is designed for efficient recovery of genomic DNA up to 60 kb in size from fresh and dried plant tissue samples. Up to 100 mg of wet tissue (or 30 mg dry tissue) can be processed in less than 1 hour. 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 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 GD2611-00 GD2611-01 GD2611-02 DNA Columns 4 50 250 2 mL collection tubes 8 100 500 Buffer P1 5 mL 40 mL 180 mL Buffer P2 1 mL 10 mL 50 mL Buffer P3 5 mL 20 mL 100 mL RNase A(20mg/mL) 30 uL 100 uL 500 uL DNA Wash Buffer 2 mL 15 mL 3 x 24 mL Elution Buffer 1.5 mL 15 mL 60 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. Dilute DNA Wash Buffer with absolute ethanol as follows and store at room temperature: GD2611-00 Add 8 mL absolute (96%-100%) ethanol GD2611-01 Add 60 mL absolute (96%-100%) ethanol GD2611-02 Add 96 mL absolute (96%-100%) ethanol to each bottle Choose the most appropriate protocol to follow. Procedures are described for each of dried and fresh (or frozen) specimens. A. Dry Specimens (Page 4) For processing ≦50 mg powdered tissue. DNA yields range from 50 μg to 100 μg per 100 mg dry tissue. B. Fresh/Frozen Specimens (Page 7) For processing ≦100 mg fresh (or frozen) tissue. DNA yield is similar to A. 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 200 μL and incubate on column at 65 ℃ for 5 min before centrifugation. No DNA DNA washed off. Dilute Wash Buffer by adding appropriate volume of absolute ethanol prior to use (page 3). 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. This product is intended for in vitro research use only. Not for use in human. This product is warranted to perform as described in its labeling and in Biomiga's literature when used in accordance with instructions. No other warranties of any kind, express or implied, including, without limitation, implied warranties of merchantability or fitness for a particular purpose, are provided by Biomiga. Biomiga's sole obligation and purchaser's exclusive remedy for breach of this warranty shall be, at the option of Biomiga, to replace the products, Biomiga shall have no liability for any direct, indirect, consequential, or incidental damage arising out of the use, the results of use, or the inability to use it product. Operating Protocol For technology support or learn more product information, please visit our website at http://www.biomiga.com/ or contact us at (858) 603-3219.Introduction
Overview
Storage and Stability
Kit Contents
Before Starting
Important
Trouble Shooting Guide
Problems
Limited Use and Warranty
- Model: GD2611-02
- Manufactured by: Biomiga