Applied Biosystems™ TaqMan™ RNase P Detection Reagents Kit

Catalog No.	Quantity
43-168-31	100 reactions

Catalog No. 43-168-31 Supplier Applied Biosystems™ Supplier No. 4316831

Description

TaqMan™ RNase P Detection Reagents Kit

The Applied Biosystems™ TaqMan™ RNase P Detection Reagents kit provides the components needed to detect and quantitate genomic copies of the human RNase P gene using the 5' nuclease assay. Human RNase P gene is a single-copy gene encoding the RNA moiety for the RNase P enzyme. This 5' nuclease assay employs TaqMan™ Universal PCR Master Mix or TaqMan™ PCR Core Reagents for polymerase chain reaction (PCR) using the provided human genomic DNA as template.

Pre-designed primers and TaqMan™ probe eliminate assay design

Easy-to-use primer and TaqMan™ probe mix minimizes assay set-up time

Rapid Assay Development Guidelines can minimize optimization time

Comprehensive Guidelines Included
Applied Biosystems™ has developed a comprehensive set of guidelines to ensure success when using Applied Biosystems™ Real-Time PCR reagents and instrumentation. These guidelines are simple and easy to follow. Many traditional variables, such as magnesium chloride concentration and the thermal cycling protocol itself, have been standardized, greatly reducing assay development time.

Normalization Built-In
All Applied Biosystems™ Real-Time PCR reagent kits contain a passive internal reference to normalize non-PCR related fluorescence fluctuations. Normalizing with a passive internal reference minimizes well-to-well variability that can result from a variety of causes, such as pipetting errors and sample evaporation. The passive reference is also essential for accurate results when multiple probes (with different reporter dyes) are combined in a single tube.

Note: See user's manual or package insert for limited label license, and trademark information. For Research Use Only. Not for use in diagnostics procedures.

Specifications

• Calibration Type: Passive Internal Reference Standards
• Content And Storage: Control Primer-Probe Set(s) & Template(s)
• Control Template: Human Genomic DNA
• Format: Tube
• GC-Rich PCR Performance: Low
• PCR Method: qPCR
• Reaction Speed: Standard
• Internal Probe Modification: TAMRA™ Quencher (3'), FAM (5')
• Label or Dye: FAM
• Product Line: TaqMan
• Product Type: Real Time PCR Reagents
• Quantity: 100 reactions
• Shipping Condition: Dry Ice
• Species: Human
• Sufficient For: 100 Reactions
• Target: RNase P
• No. of Reactions: 100 Reactions

Frequently Asked Questions (FAQs)

With the Oncomine Focus Assay, Chef-Ready Library kit, where can I find the combined barcoded AmpliSeq libraries when the run is complete?

After the completion of a run, the Ion Chef Instrument holds the barcoded libraries in the tube loaded in Position D of the Reagents cartridge. To avoid fluid loss due to evaporation, remove and cap the tube of combined barcoded libraries as soon as possible after run completion. Do not leave the tube in the instrument longer than 24 hours after the start of the run. After 24 hours from the start of the run, the instrument chiller will stop actively cooling, and the sample will be held at 27 degrees C.

The tube in Position D of the Ion AmpliSeq Chef Reagents DL8 Cartridge will contain 700 µL of combined barcoded libraries at completion of the run. The libraries are at >>100 pM (total combined library concentration) and are ready to use in template preparation. One can store unused portions of combined libraries at 4 to 8 degrees C for up to 1 month. For longer-term storage, store at -30 to -10 degrees C.

With the Oncomine Focus Assay, Chef-Ready Library kit, can I process fewer than 8 samples per Ion AmpliSeq run on the Ion Chef System?

Yes, fewer than 8 samples may be processed per run but keep in mind that a run consumes kit reagents for 8 samples regardless of the sample number.

If I process less than 8 samples using the Oncomine Focus Assay, Chef-Ready Library kit, do I need to quantitate the final combined output library?

If you are processing ≤5 samples, we recommend that you quantify your output combined library by qPCR to ensure that an optimal concentration is used in templating reactions.

If you are processing fewer than 8 samples, it is preferable to add replicates or positive control samples to the run. Otherwise, pipet 15 µL of nuclease-free water as non-template control into column 1 wells that do not contain a DNA sample.

What is the RNA input for the Oncomine Focus Assay, Chef-Ready Library kit?

Each reverse transcription reaction requires 10 ng of DNase-treated total RNA (≥1.43 ng/µL).

What is the DNA input for the Oncomine Focus Assay, Chef-Ready Library kit?

For each target amplification reaction, use 300-30,000 copies of DNA (10 ng of mammalian gDNA) from normal or FFPE tissue.

With the Oncomine BRCA Research Assay, which method do you recommend for quantifying gDNA?

We recommend two methods for gDNA quantification:

- Qubit 3.0 Fluorometer (Cat. No. Q33216) with Qubit dsDNA HS Assay Kit (Cat. Nos. Q32851, Q32854)
- qPCR using TaqMan RNase P Detection Reagents Kit (Cat. No. 4316831)

What is the purpose of the RNase P Detection Reagents Kit (Cat. No. 4316831), and what is provided in the kit?

RNase P is a single-copy gene encoding the RNA moiety for the RNase P enzyme, and it is an excellent gene to use as a reference/control for Copy Number Variation (CNV) assays. The TaqMan RNase P Detection Reagents Kit contains a 20X mix of primers and probe (FAM dye-labeled) that will detect and quantitate genomic copies of the human RNase P gene. The primers and probe are designed according to TaqMan chemistry guidelines for quantitation, and utilize the universal thermal cycling parameters. This kit is used to perform a 5' nuclease assay with TaqMan Universal PCR Master Mix (Cat. No. 4304437) or the TaqMan Core PCR Reagents Kit (Cat. No. N808-0228) with genomic, plasmid or complementary DNA (cDNA) templates.

The complete components of the TaqMan RNase P Detection Reagents Kit, Cat. No. 4316831, are: 20X RNase P Primer-Probe (FAM/TAMRA probe) Mix, 1 tube Human Genomic Control DNA, 10 ng/ul (100 ul), and the Product Insert with an overview and procedure for the use of the kit.

For Research Use Only. Not for use in diagnostic procedures.