Fluorescent Probes for Real-Time Quantitative PCR

Quality control

Every oligo is quality checked by MALDI-TOF mass spectrometry, electrospray ionization mass spectrometry (ESI-MS), capillary electrophoresis (CE), and/or polyacrylamide gel electrophoresis (PAGE).

Purification

Fully deprotected and desalted. Purified by PAGE or RP-HPLC

Length

Bases

DNA (A, C, G or T)

Backbone

Phosphodiester bond

Format

Delivered as dried-down product in opaque tubes

Turn-around time (TAT)

Turn-around time is dependent upon successful QC validation and does not include shipping time.

Storage and stability

See information on our oligos: storage recommendations

Shipment

Shipped by mail or express delivery, dry, in individual, opaque tubes

Oligonucleotide
Technical Data
Sheet

Oligonucleotides are delivered with an OligonucleotideTechnical Data Sheet, which includes oligonucleotide name, sequence, concentration, precise quantity in OD and nmols, Tm, MW, length, extinction coefficient and purification data.

Services available upon request

• Probe design services
• High-throughput screening formats (microplates, etc)
• Aliquoting
Additional services may increase turn-around time

Pricing

Please contact your local Sigma-Genosys representative

Ordering

On-line, by email or fax

What is real-time quantitative PCR?
Real-time quantitative PCR is an innovative and robust technique for quantitative analysis of gene expression, mutation detection, and SNP genotyping. Real-time quantitative PCR was developed to overcome the basic weakness of classical PCR technology: whereas PCR can indicate the presence of a particular nucleic acid in a sample after a completed PCR amplification, it cannot directly quantify the amount of amplicon.

Real-time quantitative PCR systems quantify the amount of amplified product through the detection and quantitation of a fluorescent dye. This dye, upon excitation, emits a light or signal that increases in intensity in direct proportion to the amount of amplified PCR product and, thereby, quantifies the amount of target DNA. The assays are directly performed in a single, closed tube without any purification or separation step, avoiding risks of contamination and the need for post-PCR handling.

Real-time quantitative PCR systems also enable mutation detection via melting-curve analysis, as each double-stranded DNA product has its own specific melting temperature (Tm). Melting-curve data enables the researcher to differentiate between specific PCR products and non-specific PCR products, such as primer-dimers.

Why use sequence-specific probes?
Detection by real-time quantitative PCR can be sequence-specific or non sequence specific, and the method chosen will effect the performance of your real-time quantitative PCR results. For instance, SYBR® Green 1, a non-sequence-specific reagent, can bind to any double-stranded DNA including primer-dimers and other non specific reaction products, and so may result in an over-estimation of the amount of target DNA. However, sequence-specific probes bind only to the target template and therefore achieve accurate and reliable quantification of the amount of target DNA.