Applied Biosystems™ TaqMan™ Universal Master Mix II, with UNG

Product Code.	Quantity	unitSize
10402655	1 x 5 mL	Each
10136904	1 x 1 mL	Each
10410355	2 x 5 mL	Pack of 2
10420355	5 x 5 mL	Pack of 5
10258414	10 x 5 mL	Pack of 10
10136714	1 x 50 mL	Each

Product Code. 10402655 Supplier Applied Biosystems™ Supplier No. 4440038

Description

Requires

Primers, probes, template, water

TaqMan Universal Master Mix II is a convenient mix of components (except primers, probes, template, and water) necessary to perform a real-time polymerase chain reaction (PCR).

TaqMan Universal Master Mix II is a convenient mix of components (except primers, probes, template, and water) necessary to perform a real-time polymerase chain reaction (PCR). Use TaqMan Universal Master Mix II to amplify complementary DNA (cDNA) and DNA targets for a variety of applications, including quantitation and genotyping.

This TaqMan Universal Master Mix II is supplied as a 2X concentrate and contains:
• AmpliTaq Gold DNA Polymerase, UP (Ultra Pure)
• dNTPs (with dUTP)
• ROX Passive Reference
• Uracil-N-glycosylase (UNG)
• Optimized buffer components

Note: This TaqMan Universal Master Mix II contains uracil-N-glycoslyase (UNG).

TaqMan Universal Master Mix II has been optimized for use with primers and TaqMan probes that have been designed according to Thermo Fisher Scientific guidelines. The master mix can be used with custom TaqMan assays available from our custom assay service, or with pre-optimized assays such as:
• TaqMan gene expression assays
• TaqMan microRNA assays
• TaqMan drug metabolism genotyping assays
• TaqMan SNP genotyping assays

For RNA quantitation experiments, TaqMan Universal Master Mix II is used in the second step of a two-step reverse transcription/polymerase chain reaction (RT-PCR) protocol. The cDNA template used with the master mix can be generated in a reverse transcription reaction using cDNA synthesis kits available from Thermo Fisher Scientific.

Alternative product: Try TaqMan Fast Advanced Master Mix (Cat. No. 4444963), our highest-performance, probe-based master mix. With TaqMan Fast Advanced Master Mix, we’ve taken the best of TaqMan Universal Master Mix II and added additional capabilities for your gene expression analysis.

Order Info

Guaranteed minimum shelf life is 60 days. Shipping Conditions: Wet Ice.

Specifications

• Concentration: 2X
• Content And Storage: Store at 2°C to 8°C in the dark.
  
  Guaranteed minimum shelf life is 60 days (exact expiry date printed on product and CofA).
• Detection Method: Primer-probe
• GC-Rich PCR Performance: High
• PCR Method: qPCR
• Polymerase: AmpliTaq Gold DNA Polymerase
• Reaction Speed: Standard
• For Use With (Equipment): 7300 System, 7500 System, 7900HT System,Applied Biosystems StepOnePlus™ Fast Real-Time PCR System, StepOne™, Standard Mode, StepOnePlus™, Standard Mode, ViiA™ 7 System
• Passive Reference Dye: ROX (Pre-mixed)
• Product Line: TaqMan
• Product Type: Universal Master Mix II
• Purity or Quality Grade: UP (Ultra Pure)
• Quantity: 1 x 5 mL
• Shipping Condition: Wet Ice
• For Use With (Application): Gene Expression, Genotyping, miRNA analysis
• Fidelity (vs. Taq): 2 X
• Sample Type: DNA (Genomic), cDNA
• Volume: 5 mL

Frequently Asked Questions (FAQs)

What does it mean when a miRNA target entry has been removed (obsoleted) from miRBase?

miRBase continues to re-annotate the miRNA sequences as more information becomes available. A sequence that once mapped to a given species (e.g.: human) may no longer exist, so it is removed from miRBase. Accordingly, the original assay for that sequence no longer maps to an annotated miRNA, and as a result the annotation fields on the search results page on the Thermo Fisher Scientific portal are empty. An example of this is Assay “hsa-miR-505” (Assay ID 001049). miRBase continues to re-annotate the miRNA sequences as more information becomes available. A sequence that once mapped to a given species (e.g.: human) may no longer exist, so it is removed from miRBase. Accordingly, the original assay for that sequence no longer maps to an annotated miRNA, and as a result the annotation fields on the search results page on the Thermo Fisher Scientific portal are empty. An example of this is Assay “hsa-miR-505” (Assay ID 001049).

What is the TaqMan MicroRNA Assay?

A TaqMan MicroRNA Assay has two components: an RT reaction containing a miRNA specific stem-loop reverse-transcription primer and a separate specific TaqMan miRNA Assay. The RT stem-loop primer provides the specificity for the mature miRNA target; it does not detect its precursor. The formation of a RT primer/mature miRNA-chimera extends the length of the 5’ end of the miRNA. The longer RT product provides a miRNA specific cDNA template amenable to the TaqMan assay design.

What is mechanism of the TaqMan MicroRNA Assays?

There are 2 steps involved in using the TaqMan MicroRNA Assays:

1. RT step: total RNA to cDNA using miRNA-specific RT primers from the TaqMan MicroRNA Assays and the TaqMan MicroRNA Reverse Transcription Kit (P/N 4366596, 200 reactions or 4366597, 1000 reactions)

2. PCR step: cDNA to PCR products using primers and probe from the TaqMan MicroRNA Assays together with TaqMan Universal PCR Master Mix No AmpErase UNG (P/N 4324018).

Why are miRNAs so important?

MicroRNAs are believed to play an important role in post transcriptional gene regulation. A total of ~850 unique miRNAs have been discovered so far, including ~333 human miRNAs. They are relatively abundant, ranging from a few to as many as 50,000 molecules per cell. They account for approximately 1% of the predicted genes in animals and plants. The level of individual miRNAs varies dramatically across cell type and developmental stages. These differences in miRNA level are believed to be a key indicator of miRNA activity.

Recent research has implicated miRNAs in the following: cell development, differentiation, communication and cell death; DNA methylation and chromatin modification; metabolism; human cancer development; haematopoiesis; nervous system patterning; insulin secretion.

What are miRNAs?

MicroRNAs (miRNAs) are endogenous non-coding RNAs, about 22 nucleotides in length, that play an important role in post-transcriptional gene regulation in animals and plants by targeting messenger RNA (mRNA) for degradation or translational repression. The miRNA precursors are transcribed from individual genes but are not translated into proteins. The primary transcript is processed in the nucleus to give one or more hairpin precursors that is exported to the cytoplasm. The mature miRNA is excised from the hairpin precursor. The mature miRNA is the biologically active form and regulates the expression of mRNA transcripts by binding to complementary sites on target mRNAs and inhibiting translation or, when there is perfect complementarity to the target sequence, inducing mRNA cleavage. In animals, translational repression, not transcript degradation, is the dominant miRNA mode of action.

What Applied Biosystems reagents and instruments can I use with TaqMan Gene Expression Assays?

The TaqMan Gene Expression Assays are optimized to work with one of the TaqMan qPCR Master Mixes that include TaqMan Universal PCR Master Mix, No AmpErase UNG(P/N 4324018), TaqMan Universal PCR Master Mix with AmpErase UNG (P/N 4304437), TaqMan Universal PCR Master Mix II, No AmpErase UNG (P/N 4440040), TaqMan Universal PCR Master Mix II, with AmpErase UNG (P/N 4440038) or TaqMan Gene Expression Master Mix (P/N 4369016) and with your cDNA. It is recommended to use the High Capacity cDNA Archive Kit (P/N 4322171) for converting your RNA to cDNA. TaqMan Gene Expression Assays are capable of being run on all Applied Biosystems Real-Time PCR Systems models including 7000, 7300, 7500, 7500 Fast, 7700, 7900HT, StepOne, StepOnePlus and ViiA 7 Instruments.

Can dU-containing DNA produced by amplification with dUTP be cut by restriction enzymes?

You will need to confirm with the manufacturer of the restriction enzyme you are using to find out whether or not it can recognize dUTP-containing DNA.

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