ISOLATE II miRNA Kit is specially developed for the rapid, phenol-free isolation of extremely pure and highly enriched small RNA (<200nt) including miRNA, siRNA, snRNA and piRNA, without bias (fig. 1). High quality large RNA (>200nt) such as mRNA can be easily sequentially purified from the same sample, as a separate fraction (fig. 2).
ISOLATE II miRNA Kit is optimized to deliver superior recovery from a wide range of sample sources such as mammalian tissue, cell lines, bacteria, yeast, blood and other biofluids, including challenging samples with low RNA content.
ISOLATE II miRNA Kit allows convenient processing of multiple samples in 25 minutes, without toxic chemicals such as phenol and chloroform. Isolation is based on fast spin column technology using a new generation silica. The novel separation matrix preferentially purifies small RNA from other cellular components and contaminants without laborious phenol-based phase separation and alcohol precipitation steps. In contrast, phenol-carryover is observed in preparations of small RNA using phenol-based methods and can affect performance in sensitive downstream applications.
Moreover, phenol-chloroform isolation has been reported to introduce significant bias in recovery of selected small RNA, resulting in unreliable results. ISOLATE II 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 miRNA Kit utilizes a two-column system to enrich exclusively for small RNAs, with complete diversity and no background of large RNA. Large RNA greater than 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 RNA to the second column. Once captured and enriched onto the second column, small RNA molecules are then eluted, ready-for-use.
The highly purified and enriched small RNA fraction can be used for the most sensitive applications including miRNA expression profiling and quantification using real-time PCR, RT-PCR (fig. 3), microarrays, small RNA library construction for small RNA-seq, as well as single cell assays.
|ISOLATE II Large RNA Removal Columns||25|
|ISOLATE II miRNA Columns||25|
|Collection Tubes (2ml)||50|
|Elution Tubes (1.7ml)||50|
|Lysis Buffer RX||40ml|
|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|
Certificates of Analysis for ISOLATE II miRNA Kit are grouped by catalogue number and then listed by batch number. Larger pack sizes are listed first. To locate the COA for your product, first find the catalogue number and then the required batch number.