Product Details

Bioline ISOLATE II miRNA Kit 25 Reactions (BIO-BIO-52083)

Features

ISOLATE II miRNA Kit is specially designed for the rapid and unbiased, phenol-free isolation of highly enriched small RNA
( <200nt ) with superior yield and purity, from a wide range of cells and tissues. Isolate high quality large RNA >200nt in parallel from the same sample. 

Ideal for highly sensitive applications.

  • Rapid, phenol-free workflow - unbiased small RNA isolation
  • Novel separation matrix - highest yields and purity
  • Highly specific small RNA enrichment - highest recovery of miRNA, siRNA and all other small RNA <200nt
  • Parallel large RNA purification (optional) - high quality large RNA >200nt
  • DNase I included - highly efficient digestion of genomic DNA
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Description

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.

 

Components

Reagent

25 Reactions

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

Product Manual

1

Bench Protocol Sheet

1

 

Storage & Stability

The DNase I 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.

 

Shipping conditions

Ambient temperature.

 

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Isolation of miRNA and other small RNA from

  • Mammalian tissue and cell lines
  • Bacteria and yeast
  • Biofluids (saliva, semen, urine, blood, plasma, serum)

Fully compatible with highly sensitive applications such as real-time RT-PCR, microarrays, single-cell assays and next-generation RNA sequencing (small RNA-seq).

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