PLANT DNA EXTRACTION PROTOCOL

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 INTRODUCTION


Plant materials are among the hardest for top-notch DNA extractions. The key is to appropriately set up the tissues for extraction. By and large, this includes the utilization of fluid nitrogen streak freezing followed by crushing the solidified tissue with a mortar and pestle. Fluid nitrogen is hard to deal with and it is perilous in an open research facility condition, for example, a study hall. Hence we have adjusted an extremely basic plant DNA extraction convention to utilize new tissue. We have likewise utilized tissue arranged ahead of time by desiccation. The conventions and results are introduced here. 


Extraction Protocol 


1. Weight out 0.3 g of plant tissue 


2. Spot tissue on a perfect glass slide. Hack the tissue into a glue utilizing a perfect single edge extremely sharp steel. 


3. Promptly move tissue to a 1.5 mL small scale axis tube () and (discretionary) further crush tissue with a cylinder pestle 


4. When the example is readied include 300 µL EBA, 900µl EBB, and 100 µl SDS. 


5. Vortex and brood at 65o C for 10 min. 


6. Spot tube on ice and include 410 µL cold potassium acetic acid derivation. Blend by reversal and spot tube back on the ice for 3 min. 


7. Rotator at 13,200 rpm for 15 min. (On the off chance that conceivable, utilize a refrigerated smaller scale rotator set to 4o C) 


8. Move 1 mL of the supernatant to another 1.5 mL miniaturized scale axis tube, include 540 µL of super cold supreme isopropanol, and hatch in ice for 20 min. 


9. Rotator at 10,200 rpm for 10 min. dispose of the supernatant. Wash the pellet once in 500 µL 70% ethanol and let dry 


10. Resuspend the dry pellet in 600 µL of TE. Include 60 µL 3M sodium acetic acid derivation (pH 5.2) and 360 µL super cold outright isopropanol. Hatch on ice for 20 min. 


11. Rehash Steps 9−11 twice. 


12. Resuspend the pellet in 50 µL TE and do agarose gel QC.


 

DNA

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