Small RNAs, including microRNAs (miRNAs) and small interfering RNAs (siRNAs), are important regulators of plant development and gene expression. The isolation of high-quality small RNAs is the first step in the study of its expression and function analysis.
ISOLATE II Plant miRNA Kit is based on the latest in rapid spin column technology using a novel RNA silica as the separation matrix for the isolation of high quality and highly enriched small RNA species (<200nt) including miRNA, siRNA and tRNA. Small RNA are preferentially purified from other cellular components without the use of phenol or chloroform. High quality large RNA (>200nt) can be sequentially purified from the same sample. The kit is compatible with a wide range of plant cells and tissues (Fig. 1).
ISOLATE II Plant miRNA Kit has been developed to overcome the bias observed with small RNA isolation using phenol-based techniques, due to its proprietary small RNA separation and enrichment technology, as well as highly optimized chemistry.
ISOLATE II Plant miRNA kit allows convenient processing of multiple samples in 25 minutes, utilizing a two-column system to enrich exclusively for small RNAs. RNAs greater than approximately 200 nucleotides are bound to the first column and ethanol is then added to the flow-through in order to increase the binding affinity of small RNAs to the second column. Once bound to the second column, small RNA molecules are then eluted, ready-for-use in a wide variety of downstream applications.
Selected plant samples isolated with the ISOLATE II Plant miRNA Kit
|Tobacco (Nicotiana tabacum)||Leaf, stem, root|
|Cotton (Gossypium)||Leaf, cotton|
|Soy bean||Leaf, stem, root|
|Potato (Solanum tuberosum)||Leaf, tuber|
|Corn/Maize (Zea mays)||Leaf, corn|
|Rice (Oryza sativa)||Leaf|
|Thale cress (Arabidopsis thaliana)||Leaf, stem|
|Tomato (Lycopersicon esculentum)||Leaf|
|Strawberry||Leaf, fruit, flower|
|Apple (Malus sp.)||Leaf, flower, pollen|
|Plum (Prunus sp.)||Leaf, fruit|
|Peach (Prunus persica)||Leaf|
|Pear (Pyrus sp.)||Leaf|
|Grape vine (Vitis sp.)||Leaf, grape, skin|
|Pepper (Capsicum annuum )||Leaf, seed|
|Palm (Arecaceae) (Elaeis guineensis)||Leaf|
|Eastern white Red Cedar||Leaf|
|Pine needle (Pinaceae)||Needle|
|Rose gum (Eucalyptus grandis)||Root|
|ISOLATE II BioFilters||25|
|ISOLATE II Large RNA Removal Columns||25|
|ISOLATE II miRNA Columns||25|
|Collection Tubes (2ml)||50|
|Collection Tubes (2ml)||25|
|Elution Tubes (1.7ml)||50|
|Lysis Buffer RPX||30ml|
|Wash Buffer W1||38ml|
|DNase I Solution (RNase-free)||0.8ml|
|DNase I Reaction Buffer DRB||6ml|
|RNA Elution Buffer||6ml|
|Bench Protocol Sheet||1|
The DNase I Solution should be stored at -20°C upon arrival. All other components should be stored dry and at room temperature.
When stored under the recommended conditions and handled correctly, full activity of reagents is retained until the expiry date indicated on the outer box label.