/ USBio Product-Datasheet M3949-97

Biomol GmbH
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22769 HAMBURG
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Specifications

Catalog #M3949-97
IntroductionMicroRNAs (miRNAs) are endogenous regulators of gene expression that are encoded in the genomes of plants, animals and viruses, then processed into ~19-23 nt, single-stranded molecules that become incorporated into the RNA-induced silencing complex (RISC). RISC mediates down-regulation of gene expression through translational inhibition, transcript cleavage, or both. There is emerging evidence that some microRNAs can function as oncogenes or tumor suppressors. Recent study showed that microRNA-10b (miR-10b) is highly expressed in metastatic breast cancer cells and positively regulates cell migration and invasion. Designed for use with MicroRNA qRT-PCR primer sets, the miR-10b One-Step qRT-PCR Detection Kit provides a one-step, simple, robust, inexpensive assay for quantitative analysis of mir-10b expression from total RNA samples, or small RNAs enriched samples. MicroRNA qRT-PCR primer sets were uniquely designed and optimized for detecting mature miRNA with high specificity and sensitivity at low PCR annealing temperature. The reverse PCR primer is also served as RT primer for reverse transcription. This kit contains an enzyme mix, a 2X-concentrated qRT-PCR reaction mixture and a miRNA primer set that contain all the reagents (except templates) needed for running qRT-PCR assay. The passive reference dye ROX is included in a separate tube to make this kit adaptable for many real-time QPCR platforms. Total RNA from metastatic breast cancer cell line MDA-MB-231 is supplied with the kit as positive control.
Features Convenient - All reaction components are supplied for quick and easy set up Save time One-step qRT-PCR procedure reduces setup time and liquid handling steps Wide dynamic range: good linearity and excellent PCR efficiency over a 6 logs of dynamic range High Sensitivity - detect miRNA expression in as low as 2.5pg total RNA. Flexible Compatible with most of the real-time PCR instruments.
Applications Mir-10b detection and quantification Breast cancer metastasis studies
DescriptionComponents in this kit are prepared with pure chemicals according to our proprietary technology. Mir-10b One-Step qRT-PCR Detection Kit provides a one-step, simple, robust, inexpensive assay for detection and quantitative analysis of mir-10b expression from total RNA samples, or small RNAs enriched samples. Quality Control 1 kit of this lot has been tested for amplifying miR-10b from total RNA of metastatic breast cancer cell line MDA-MB-231 over a 6 logs of dynamic range using Stratagene's Mx3005P as a real time PCR instrument. Good linearity and great PCR efficiency is observed and consistent with the previous lot.
ComponentsReagents are sufficient for 200 assays.
Item Amount
1. MicroRNA 2X qRT-PCR Reaction Mixture, 1.25ml x 2
2. MicroRNA qRT-PCR Enzyme Mix*, 100ul
3. ROX Reference Dye, 100ul
4. miR-10b qRT-PCR Primer Set (25X) 200ul
5. MDA-MB-231 Total RNA (25ng/ul) 100ul
6. Nuclease-Free PCR Grade Water 3ml
* microRNA qRT-PCR enzyme mix contains Reverse Transcriptase, RNase Inhibitor, and Hotstart Taq DNA Polymerase.
Storage and Stability Upon receipt, store all components at -20°C in a constant temperature freezer. Avoid repeated freeze/thaw cycles. When stored under these conditions this kit is stable for one year after ship date. The dye in MicroRNA 2x qRT-PCR Mixture and the ROX reference dye are light sensitive and should be kept away from light whenever possible.
Important NoteThis product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications without the expressed written authorization of United States Biological.


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