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miScript miRNA PCR Array Service

For miRNA profiling of pathway- or disease-focused miRNAs or entire miRNomes

  • Sensitive, specific miRNA profiling
  • Profiling pathway- or disease-focused miRNAs or the whole miRNome
  • Outstanding data quality
  • Stringent quality control

The miScript miRNA PCR Array Service uses the miScript PCR System to profile the expression of your miRNAs of interest. miScript miRNA PCR Arrays are used for pathway-focused, disease-focused, or whole miRNome expression profiling.

Cat No./ID: Inquir
miScript miRNA PCR Array Service
miRNA profiling service for pathway- or disease-specific miRNAs or whole miRNomes
The miScript miRNA PCR Array Service is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
Principle
The miScript miRNA PCR Array Service uses your samples to perform sensitive, specific miRNA profiling using SYBR® Green-based real-time PCR. The Service covers all the steps involved in isolation of total RNA (including miRNA), conversion of RNA to cDNA, real-time PCR detection of miRNAs, and data analysis. 
miScript miRNA PCR Arrays use cutting-edge technology that integrates universal tailing and reverse transcription with miRNA-specific PCR. Profiling is highly sensitive - as few as 10 miRNA copies can be detected. In addition, high primer specificity is assured with efficient discrimination between closely related isoforms, even when the miRNAs only differ by a single base. Validated procedures and optimized processes yield the highest data quality and fastest turnaround time.
Procedure

The miScript miRNA PCR Array Service includes:

Total RNA (including miRNA) purification
Purification of total RNA from cells, tissues, blood, formalin-fixed paraffin-embedded (FFPE) samples, laser capture microdissection (LCM) samples, fine needle aspiration biopsies (FNAB), and other small samples if required 
Reverse transcription using the miScript II RT Kit
miRNA profiling using a pathway-focused, disease-focused, or whole miRNome miScript miRNA PCR Array
Data analysis
Sample processing

For instructions regarding sample submission and shipping, contact QIAGEN Technical Services. Samples accepted are total RNA, cells, or tissue. An RNA Isolation Service is provided if required.

Before shipping:

Obtain a sample submission form from QIAGEN Technical Services.
Complete the form and include it with the sample shipment.
Include the amount of material and its storage conditions.
Sign and date the form.
Ship according to the instructions for your sample type.
Submitting cells or tissue samples

All sample collection should be carried out in a timely manner in an RNase free environment. Pay close attention to the minimum sample amounts required. If you have less than these minimum amounts, or your previously isolated samples have been stored differently then we recommend, contact QIAGEN Technical Services before submitting your samples.

Cells:

Provide at least 1 million (106) cells for each sample.
Specify the number of cells on the order form.
Snap freeze the cell pellets in liquid nitrogen.
Alternatively, provide homogenized cell lysates (see the miRNeasy Mini Handbook for details of lysis using QIAzol Lysis Reagent)
Store samples frozen at -80 ºC, and ship on dry ice.

Tissue:

If possible, provide at least 50 mg of tissue for each sample.
If possible, specify the weight of the tissue on the order form.
Upon collection, stabilize samples in RNAlater RNA Stabilization Reagent or AllProtect Tissue Reagent overnight.
After overnight incubation, remove excess reagent before freezing.
Snap freeze tissues in liquid nitrogen.
Store samples frozen at -80 ºC, and ship on dry ice.

FFPE samples:

Consult QIAGEN Technical Services before isolating and submitting samples, and preferably before sample collection.
For slide-mounted tissue sections, submit at least 4 slides with at least 5- to 10-micron thick sections. More slides may be requested from certain tissue sources, if available.
For freshly cut sections of whole FFPE blocks, submit three to four 20-micron thick sections.
Specify fixation procedure and age of samples.
Store and ship at ambient temperature.

LCM samples:

Consult QIAGEN Technical Services before isolating and submitting samples.
Store samples frozen at -80 ºC, and ship on dry ice.

FNAB and other small samples:
Consult QIAGEN Technical Services before isolating and submitting samples.

Other sample types:

Consult QIAGEN Technical Services before isolating and submitting samples.

Submitting purified RNA

RNA samples are very sensitive to RNase digestion. Wear gloves and maintain an RNase-free work area while purifying RNA.

Recommended RNA purification methods
Sample typeMethod
Cultured cells miRNeasy Kit or RNeasy Plus Kit (used with QIAGEN Supplementary Protocol: Purification of miRNA from animal cells using the RNeasy Plus Mini Kit and RNeasy MinElute Cleanup Kit) or RNeasy Plus Micro Kit
Tissue miRNeasy Kit
FNAB miRNeasy Kit
FFPE samples miRNeasy FFPE Kit
LCM samples miRNeasy FFPE Kit for samples from FFPE sections; miRNeasy Kit for samples from cryosections
Small samples yielding less than 100 ng total RNA miRNeasy Kit
Whole blood Before RNA preparation, red blood cells must be removed from whole blood samples using either selective lysis (e.g., as described in the QIAamp RNA Blood Mini Handbook) or a density gradient centrifugation medium (e.g., Lymphoprep, Greiner Bio-One [cat. no. 1031966]). RNA should then be purified from the white blood cell fraction using the miRNeasy Mini Kit. Alternative methods of RNA purification include use of the PAXgene Blood RNA Kit.
Total RNA isolated using a phenol-based method Clean up the RNA using an miRNeasy Kit
Other sample types Contact QIAGEN Technical Services

For best results from the PCR Array, all RNA samples should be suspended in the RNase-free water provided with the RNA Isolation kit. Do not use DEPC-treated water.

Note: Total RNA preps are highly suitable for use with the miScript miRNA PCR Array Service. It is not necessary to provide small RNA-enriched preps.

Quality control

We recommend verifying the quality of purified RNA using UV spectrophotometry. Please submit the results to the miScript miRNA PCR Array Service for evaluation.
To assess RNA quality by examining ribosomal RNA band integrity, electrophorese a fraction of each RNA sample on a denaturing agarose gel or on the QIAxcel.

Note: Due to the fragmentation associated with formalin fixation and paraffin embedding, RNA isolated from FFPE blocks and slides is always degraded, and the extent of degradation varies among samples. Therefore, while the integrity of RNA isolated from FFPE blocks or slides may be determined by the methods described here, it may only be useful for the purposes of comparing samples.

Note: RNA yields from some sources, such as serum or plasma samples, LCM samples, and FNAB samples are too low to enable quality control by these methods.
Data delivery

Data analysis is performed using QIAGEN data analysis software. Data in an electronic form can be downloaded from a secure FTP Website or delivered via email. A complete data and experiment report includes:

Raw CT values
ΔΔCT Analysis
Excel spreadsheet (data analysis using pair-wise comparisons, including raw data, tabular results as well as Scatter, 3D, and Volcano plots)
Applications

The miScript miRNA PCR Array Service enables:

Disease- or pathway-focused miRNA profiling
Whole miRNome profiling
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