Can any one suggest me a list of Genomic DNA extraction kits available to extract Genomic DNA from rice seed and rice plant leaf tissue. Rice seed is rich in amylose content. Especially those of pigmented varieties that are rich in flavanoids and anthocyanadins. Can anyone please suggest commercially available kit so that we could proceed successfully with our experiments.
Regards
Sakthi kumaran
Hi , I always used Qiagen kits for DNA isolation while working with plants and specifically with rice plant. kindly check the link:
https://www.qiagen.com/us/shop/sample-technologies/dna/genomic-dna/dneasy-plant-mini-kit/#orderinginformation
use this protocol :
Zaidani et al., 2005.
• Leaf sample of 300 mg each of present genotypes were weighed.
• All genotypes were separately crushed in the liquid nitrogen by the use mortar and pestle. The powdered form of sample was collected in micro centrifuge tubes.
• Pre-warmed extraction buffer (1ml) containing 200 µl β-mercaptoethanol (added freshly) was added in to each micro centrifuge tube (2 ml) and mix it well by vortexing.
• The tubes were incubated at 65oC for 1 hr with frequent swirling.
• An equal volume of Chloroform: Isoamyl alcohol (24:1) was added in to each micro centrifuge tube and centrifuged at 10,000 rpm at 4oC for 10 min.
• The supernatants were pooled out from the each micro centrifuge tube and taken in another micro centrifuge tube. Again Chloroform: isoamyl alcohol (24:1) was added and again centrifuged. This step was repeated twice.
• The DNA was precipitated by the use of absolute alcohol (2/3 vol) and kept for 12 h.
• The precipitated DNA was washed with 70% alcohol and air-dried.
• 100µl of TE buffer was added to the air-dried pellet and dissolved. To this 2µl of RNase was added and incubated initially at 37ºC for 50 min and later at 65ºC for 10 min.
• The DNA sample was then analyzed for its purity by Spectrophotometric Analysis.
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