<D2411> Plant DNA DS Mini Kit DS植物DNA提取試劑盒
DS植物DNA提取試劑盒
Plant DNA DS Mini Kit
產品編號 | 產品名稱 | 包裝規格 | 目錄價 |
D2411-00 | DS植物DNA提取試劑盒 | 5T | 160 |
D2411-01 | DS植物DNA提取試劑盒 | 50T | 1250 |
產品介紹
The E.Z.N.A. Plant DNA DS Kit is designed for the efficient recovery of genomic DNA up to 30 kb in size from fresh, frozen or dried plant tissue samples rich in polysaccharides, polyphenols, or those with a lower DNA content. Up to 50 mg wet tissue (or 15 mg dry tissue) can be processed in less than 1 hour. These systems combine 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 enzyme digestion and hybridization applications.
These procedures rely on the well-established properties of the cationic detergent, cetyltrimethyl ammonium bromide (CTAB), in conjunction with the unique binding system to increase yields and provide high-quality DNA. The system eliminates the need for chloroform extractions traditionally associated with CTAB-based lysis methods. Samples are homogenized and lysed in a high salt buffer containing CTAB and with binding conditions optimized, DNA is purified using a DNA mini column. Salts, proteins, and other contaminants are removed to yield high-quality genomic DNA.
參數
FEATURES | SPECIFICATIONS |
Starting Amount | Up to 50 mg wet tissue |
Starting Material | Fresh, frozen, or dried plant tissue samples rich in polysaccharides, polyphenols, or those having a lower DNA content |
Elution Volume | 50–100 μL |
Technology | HiBind? DNA Mini Column |
Processing Mode | Manual |
Throughput | 1–24 |
Note | CTAB lysis |
組分
Product | D2411-00 | D2411-01 | D2411-02 |
Purifications | 5 | 50 | 200 |
HiBind? DNA Mini Columns | 5 | 50 | 200 |
Homogenizer Mini Columns | 5 | 50 | 200 |
2 mL Collection Tubes | 10 | 100 | 400 |
CSPL Buffer | 5 mL | 40 mL | 160 mL |
RBB Buffer | 5 mL | 30 mL | 120 mL |
XP2 Binding Buffer | 5 mL | 30 mL | 2 × 60 mL |
HBC Buffer | 5 mL | 25 mL | 80 mL |
DNA Wash Buffer | 2.5 mL | 25 mL | 100 mL |
Elution Buffer | 2 mL | 30 mL | 30 mL |
Proteinase K Solution | 150 μL | 300 μL | 4.4 mL |
RNase A (25 mg/mL) | 30 μL | 300 μL | 1.2 mL |
User Manual | ? | ? | ? |
流程
DNA Extraction and Purification from Wet/Frozen/Dry Tissue Samples
Prepare 10-50 mg wet/frozen tissue or 2-10 mg dry tissue in a 1.5 or 2 mL microcentrifuge tube/vial (not provided). See reverse for sample disruption techniques. For best results, use a commercial homogenizer if available.
Add 700 μL CSPL Buffer and 20 μL Proteinase K Solution. Vortex vigorously to mix. Make sure to disperse all clumps.
Incubate at 65°C for 30 minutes. Centrifuge at 12,000g for 3 minutes.
Insert a Homogenizer Mini Column into a 2 mL Collection Tube.
Transfer 550 μL cleared supernatant to the Homogenizer Mini Column. Centrifuge at 12,000g for 1 minute.
Transfer the filtrate to a new 2 mL microcentrifuge tube (not provided). Add 5 μL RNase A. Let sit at room temperature for 5 minutes.
Add 525 μL RBB Buffer and 525 μL XP2 Binding Buffer. Vortex to mix thoroughly.
Insert a HiBind? DNA Mini Column into a 2 mL Collection Tube.
Transfer 750 μL lysate from Step 6 to the HiBind? DNA Mini Column. Centrifuge at 12,000g for 1 minute. Discard the filtrate and reuse the collection tube.
Repeat Step 8 to transfer the remaining lysate.
Add 500 μL HBC Buffer diluted with 100% isopropanol (see the bottle for instructions). Centrifuge at 12,000g for 1 minute. Discard the filtrate and reuse collection tube.
Add 700 μL DNA Wash Buffer diluted with 100% ethanol (see the bottle for instructions). Centrifuge at 12,000g for 1 minute. Discard the filtrate and reuse the collection tube.
Repeat Step 12 for a second DNA Wash Buffer Step.
Centrifuge the empty HiBind? DNA Mini Column at 12,000g for 2 minutes to dry the column. This step is critical for removal of trace ethanol that may interfere with downstream applications.
Transfer the HiBind? DNA Mini Column to a clean 1.5 mL microcentrifuge tube (not provided).
Add 50-100 μL Elution Buffer heated to 65°C directly to the center of the column membrane. Let sit at room temperature for 1 minute. Centrifuge at 12,000g for 1 minute.
Transfer the filtrate to the center of the HiBind? DNA Mini Column membrane. Let sit at room temperature for 1 minute. Centrifuge at 12,000g for 1 minute.
Store filtrate containing DNA at -20°C.
Disruption of Plant Tissue
*Grind samples with pestle
A) Dry Specimens
Drying allows storage of field specimens for prolonged periods of time prior to processing. Samples can be dried overnight in a 45°C oven, powdered, and stored dry at room temperature. To prepare dried samples, place ~15 mg dried tissue into a microcentrifuge tube (1.5 mL tubes are recommended) and grind using a pellet pestle. A fine powder will ensure optimal DNA extraction and yield.
B) Fresh/Frozen Specimens
Due to the tremendous variation in water and polysaccharide content of plants, sample size should be limited to ~30 mg for first time users. It is very important to not overload the HiBind? DNA Mini Column. Too much starting material will decrease the yield and purity due to inefficient lysis. We recommend starting with 30 mg tissue. If results obtained are satisfactory, then increase amount of starting material. Best results are obtained with young leaves or needles.
To prepare samples, collect tissue in a 1.5 mL or 2 mL microcentrifuge tube and dip the tube in liquid nitrogen with a pair of tweezers to fill the tube. Grind the tissue using disposable Kontes pellet pestles. Alternatively, allow the liquid nitrogen to evaporate and store the samples at -70°C for later use. Transfer the ground sample into a 1.5 mL microcentrifuge tube.
Note: Do not allow the sample to thaw during handling and weighing. To prevent the sample from thawing, keep the samples on a bed of dry ice.
*Disrupt Samples With Commercial Homogenizers
Fresh, frozen, and dried plant tissue can be effectively disrupted and homogenized by rapid agitation in the presence of beads.
Add two 3-4 mm stainless steel beads or ceramic beads to each vial.
Add 700 μL CSPL Buffer and 20 μL Proteinase K Solution to each sample.
Close the individual vial.
Place the racks or plates into the clamps of the homogenizer.
Homogenize for 60-90 seconds at 30 Hz. Tissue samples are disrupted and simultaneously homogenized with the shearing and crushing action of the beads.
Continue to Step 3 of the DNA Extraction and Purification Protocol on the reverse page.
公司名稱：上海諾寧生物科技有限公司
聯系Q Q：306458042（經銷商）392324729（終端用戶） 
聯系微信：NoninBioTech（經銷商）shnoninbio（終端用戶）
聯系地址：上海市閔行區虹梅南路2588號綠亮科創園A531
聯系電話：021-61241614 18616364863
E-mail：noninbio@163.com
