Navigation Links
DNA Isolation From 300 to 450 mg Dried or 600 to 1,200 mg Fresh Leaf Tissue


AquaPure Genomic DNA Isolation Kit
Catalog #732-6340
Expected yield range: 90900 g DNA


Method
Cell Lysis
1. Add 300-450 mg dried tissue (finely ground) or 6001,200 mg frozen tissue (which may be finely ground with a mortar and pestle in liquid nitrogen), or 6001,200 mg fresh tissue to a 50 ml tube. Work quickly and keep tissue cold to minimize DNase activity. Note: it may be necessary to vary the amount of starting material depending upon the species, age, tissue preparation, and genome size.

2. Add 18 ml cell lysis solution to the leaf tissue. For dried tissue, cap the tubes and vortex 13 sec to wet the tissue. For unground tissue, homogenize using 3050 strokes with a tube pestle.

3. Incubate cell lysate at 65C for 60 min. After 30 and 60 min, invert capped tube 10 times.

RNase Treatment (Optional)
1. Add 90 l RNase A solution to the cell lysate.

2. Mix the sample by inverting the capped tube 25 times and incubate at 37C for 15 min.

Protein Precipitation
1. Cool sample to room temperature.

2. Add 6 ml protein precipitation solution to the lysate.

3. Cap the samples and vortex vigorously at high speed for 20 sec to mix the protein precipitation solution uniformly with the cell lysate. Alternatively, invert a rack containing the samples 150 times (approximately 2 min) to mix the protein p recipitation solution uniformly with the cell lysate. For species with high polysaccharide content, it may be necessary to incubate sample on ice for 515 min.

4. Centrifuge at 3,000 x g for 10 min. The precipitated proteins should form a tight, green pellet. The supernatant may range in appearance from a transparent brown to green, depending on the sample. If the pellet is not tight, repeat step 3, incubate on ice for 5 min, then repeat step 4.

DNA Precipitation
1. Leaving behind the precipitated protein pellet, pour the supernatant containing the DNA into a 50 ml centrifuge tube that contains 18 ml 100% isopropanol (2-propanol).

2. Mix the sample by inverting the capped tube gently 50 times.

3. Centrifuge at 3,000 x g for 5 min. The DNA will be visible as a pellet that ranges in color from off-white to dark green, depending on particulates that cosediment with the DNA.

4. Pour off supernatant and drain tube briefly on clean absorbent paper. Add 18 ml 70% ethanol and invert the capped tube several times to wash the DNA pellet.

5. Centrifuge at 3,000 x g for 5 min. Carefully pour off the ethanol. Pellet may be loose, so pour slowly and watch the pellet to ensure that it stays in the tube.

6. Invert and drain the tube on clean absorbent paper and allow to air-dry 15 min.

DNA Hydration
1. Add 1 ml DNA hydration solution.

2. Rehydrate DNA by incubating sample 1 hr at 65C, or a l t e rn a t i v e l y, overnight at room temperature. Tap tube periodically to aid in dispersing the DNA. Transfer DNA sample to a clean 1.5 ml tube for storage.

3. If particulates are present in the rehydrated DNA sample, centrifuge at 13,00016,000 x g for 510 min and transfer supernatant into a clean 1.5 ml tube. Store DNA at 28C.


back to top
'"/>

Source:


Page: All 1 2

Related biology technology :

1. Fast and Easy Isolation of PCR-Ready Genomic DNA from Whole Blood
2. High-Throughput Isolation of Total RNA
3. Simple Isolation of RNA from Tissue and Cultured Cells
4. Fast Isolation of Total RNA from Small Numbers of Cells
5. Comparison of Growth Techniques and Media for the Purpose of Plasmid Isolation from E. coli Using the Eppendorf Perfectprep Plasmid Mini Kit
6. Isolation of Microorganisms
7. Isolation of Genomic DNA from Saliva Using the Perfect gDNA Blood Mini Kit
8. Mouse Tail Genomic DNA Isolation Protocol(1)
9. Genomic DNA Isolation Protocol(1,2,3)
10. Basic Plasmid DNA Isolation Protocol
11. Protocol for RNA Isolation Using TRIzol Reagent with Phase Lock Gel-Heavy
Post Your Comments:
*Name:
*Comment:
*Email:
TAG: DNA Isolation From Dried Fresh Leaf Tissue

(Date:2/27/2015)... , Feb. 27, 2015  Steep Hill, the industry ... and internationally, intends to open a full service medical ... Mexico , bringing advanced scientific tools and methodology ... Steep Hill is currently the only laboratory licensed ... potency and contaminant testing in order to meet the ...
(Date:2/26/2015)... 2015 /PRNewswire/ - SQI Diagnostics Inc. ("SQI" or the ... that develops and commercializes proprietary technologies and products for ... results for the fiscal first quarter ended December  31, ... Canadian dollars (CAD), unless otherwise stated. ... business from our existing customers and add to our ...
(Date:2/26/2015)... (PRWEB) February 26, 2015 The Movement ... demonstration to support life extension technologies and awareness. The ... Universal Coordinated Time, on March 21st 2015, in a ... , Eric Schulke, Founder of MILE, says, “Our ... with urgency. The Movement for Indefinite Life Extension is ...
(Date:2/26/2015)... February 26, 2015 The Pittcon ... at the Ernest N. Morial Convention Center in ... as of February 19, 2015) displaying products and ... academic, and government labs. The Exposition will offer ... not limited to, analytical chemistry; drug discovery; nanotechnology; ...
Breaking Biology Technology:Steep Hill First to Receive Provisional Approval to Establish Laboratory Services for Cannabis Testing in State of New Mexico 2SQI Diagnostics Reports First Quarter Financial Results 2SQI Diagnostics Reports First Quarter Financial Results 3SQI Diagnostics Reports First Quarter Financial Results 4SQI Diagnostics Reports First Quarter Financial Results 5SQI Diagnostics Reports First Quarter Financial Results 6Indefinite Life Extension Activists Organize Online Demonstration 2Indefinite Life Extension Activists Organize Online Demonstration 3Pittcon 2015 Announces Exposition Highlights 2
... at M. D. Anderson Cancer Center has been charged with providing ... , ... HOUSTON (Business Wire EON) July 1, 2008 ... to document activities within the laboratory workflow at its siRNA facility, ...
... 1 IBT Laboratories, a leading,global clinical-diagnostics ... assays, announced that Maureen Loftus has joined,the ... According to IBT,s,founder and Chief Executive Officer, ... leadership skills and experience, garnered from more ...
... PRINCETON, N.J., July 1 Pharmasset, Inc.,(Nasdaq: VRUS ... reconstituted its equity indexes on June 27., Pharmasset,s ... for,one year, means automatic inclusion in the small-cap Russell ... style indexes. Russell indexes,are widely used by investment managers ...
Cached Biology Technology: M. D. Anderson Automates Laboratory Workflow with Stone Bond Technologies : EE-LIMS™ from Stone Bond Technologies Will Document Activities within the Laboratory Workflow at the siRNA Facility, Increasing Throughput, While Reducing Risks 2IBT Laboratories Names Maureen Loftus President and Chief Operating Officer 2IBT Laboratories Names Maureen Loftus President and Chief Operating Officer 3Pharmasset Joins Russell 3000 Index 2
(Date:2/5/2015)... TRIANGLE PARK, North Carolina , 5. ... sein 35. Jahr als spezialisiertes Logistikunternehmen und ... Position als einzige klinische Logistikfirma (Clinical Logistics ... Die Kernbotschaft der neuen Kampagne lautet ... und Lieferungen. Foto: http://photos.prnewswire.com/prnh/20150205/173753 ...
(Date:2/5/2015)... 2015   Epic Sciences , a precision diagnostics company ... Murali Prahalad , Ph.D., president and CEO, is scheduled ... (PMWC) 2015: Silicon Valley, which is taking place at the ... on January 26-28, 2015. Dr. Prahalad ... Biomarkers." Last year, Epic Sciences was a finalist in the ...
(Date:2/5/2015)... Jan. 27, 2015   Marvin Test ... deployed, innovative test solutions for military, aerospace, ... of its TS-323 GENASYS Test Platform ... (LMSSC). GENASYS is a high-performance PXI-based system ... applications that require performance functional testing. GENASYS ...
Breaking Biology News(10 mins):Marken startet Kampagne mit Patienten im Mittelpunkt 2Marken startet Kampagne mit Patienten im Mittelpunkt 3Epic Sciences to Present at Personalized Medicine World Conference 2015 on the Use of Circulating Tumor Cells as Cancer Biomarkers 2Marvin Test Solutions' GENASYS Platform Advances Mission-Critical Aerospace Production Test 2Marvin Test Solutions' GENASYS Platform Advances Mission-Critical Aerospace Production Test 3Marvin Test Solutions' GENASYS Platform Advances Mission-Critical Aerospace Production Test 4
... HOUSTON (July 9, 2012) -- What,s 100 times stronger ... like a twig by a tiny air bubble? The answer ... Rice University scientists details exactly how the much-studied nanomaterials snap ... find that the old saying ,I will break but not ...
... During a meeting in Banaue, The Philippines, scientists from ... UK, Bulgaria and Spain raised several concerns on the ... The meeting was organized within the framework of the ... multiple risks for rice ecosystems arising from various aspects ...
... Researchers at the RUB and from Berkeley have used ... Journal of the American Chemical Society , they report ... the resulting metal-peptide compounds. "With this work, we have ... medicines" says Prof. Raphael Stoll from the Faculty of ...
Cached Biology News:Tiny bubbles snap carbon nanotubes like twigs 2Tiny bubbles snap carbon nanotubes like twigs 3Sustainability of rice landscapes in South East Asia threatened 2A new avenue to better medicines: Metal-peptide complexes 2
Mycoplasma and virus tested...
Bethyl Laboratories Ready-To-Use IHC Bluing Reagent is used following IHC hematoxylin counterstaining with immunohistochemical procedures. Shelf-Life: 1 year from date of receipt...
Edited by Benjamin Lewin (1997) • This book gives an integrated account of the structure and function of genes in both prokaryotic organisms and is thoroughly up to date with the latest thinking...
... TOPO Cloning is ... available for cloning PCR ... and Zero Blunt TOPO ... and activated with topoisomerase ...
Biology Products: