I suggest to extract twice with TRIZol reagent, following manufacturer's instructions. I mean extract and resuspend your RNA in DEPC-water, and then repeat the RNA-extraction with TRIZol reagent, and finally resuspend your RNA in DEPC-water. After that procedure quantify your RNA, and you might see and increase in your 260/280 and 260/230 -ratio, that means an improvement in RNA quality. Albeit, you are gonna lose some amount of your RNA, but its better to work for qPCR.
Regards
RNA contaminants like sweat and others may also reduce the quality of our petal rose RNA.. Ensure gloves are worn and all apparatus properly sterilized, mop all bench, glass and plastic surfaces with RNase before use. Depending on your extraction buffer, you may filter and add higher volume of B-Mecaptoethanol to extraction buffer before use . Resuspend in 20ul of DEPC - water..
To date, no commercial kit has been developed for the isolation of high-quality RNA from tissues with these characteristics, especially for fruit. The common protocols for RNA isolation are tedious and usually result in poor yields when applied to recalcitrant plant tissues. Here an efficient RNA isolation protocol based on cetyltrimethylammonium bromide (CTAB) and two successive precipitations with 10 M lithium chloride (LiCl) was developed specifically for loquat fruits, but it was proved to work efficiently in other tissues of loquat and woody plants. The RNA isolated by this improved protocol was not only of high purity and integrity (A260/A280 ratios ranged from 1.90 to 2.04 and A260/A230 ratios were>2.0) but also of high yield (up to 720 μg on average [coefficient of variation=21%] total RNA per gram fresh tissue). The protocol was tested on loquat fruit (different stages of development, postharvest, ripening, and bruising), leaf, root, flower, stem, and bud; quince fruit and root; grapevine cells in liquid culture; and rose petals. The RNA obtained with this method is amenable to enzymatic treatments and can be efficiently applied for research on gene characterization, expression, and function.
i am also having a lot of trouble in RNA extraction from Rose tissues (leaves, flower buds, etc). I have used pBiozol, but still i haven't been successive in having a brighter 28s band in gel. I have repeated about 30 times, with different criteria, but all in vain. Please help me :`(
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