Real-Time PCR
Real-time/Quantitative (q)-PCR is a powerful advancement of the basic PCR technology. Through the use of appropriate fluorescent detection strategies in conjunction with proper instrumentation, the starting amount of nucleic acid in the reaction can be quantified. Quantification is achieved by measuring the increase in fluorescence during the exponential phase of PCR. Quantitative PCR has a sensitivity five orders of magnitude better than the best blotting procedures and a dynamic range of 10 orders of magnitude! These features made the work of turning PCR into a quantitative tool worthwhile which is reflected by the continuing increase of popularity for different kinds of real time/quantitative PCR Systems.
Applications of q-PCR are found in many key areas including:
- Gene Therapy Distribution and Expression Assays
- Drug Response Analysis
- Cell Bank Copy Number Analysis
- Mutation Analysis/SNP Analysis
- Expression System Development
- Residual DNA Analysis
- Detection of Pathogens
- GMO Analysis
- Human and Veterinary Diagnostics
- Blood products quality control
metabion can supply you probes for every real-time PCR instrument available in the market. No matter if you work with any of ABI's Prism 7XXX line machines, Corbett's Rotogene, BioRad's I-Cycler, Cepheid's Smart Cycler, MJ Research's Opticons, or Roche's LightCycler® - we have a solution for you.
able 1: Reporter/3'Quencher combinations
For a list of compatible probes and equipment (.pdf)
| 3' Quencher |
| |
TAMRA |
BHQ-1 |
BHQ-2 |
BHQ-3 |
Eclipse |
Dabcyl |
|
|
|
| Abs range (nm) |
470-560 |
480-580 |
550-650 |
620-730 |
390-625 |
380-550 |
|
|
|
| Abs max (nm) |
544 |
535 |
579 |
672 |
522 |
453 |
|
|
|
| 5' Reporter |
Pricing Level |
Abs (nm)* |
Em (nm)* |
Color |
| LC® CYAN500 |
- |
3 |
- |
- |
3 |
3 |
450 |
500 |
|
| Fluo |
1 |
1 |
1 |
- |
1 |
1 |
495 |
520 |
|
| 6-FAM |
1 |
1 |
1 |
- |
1 |
1 |
495 |
520 |
|
| Rhodamine Green |
- |
3 |
- |
- |
3 |
3 |
503 |
528 |
|
| TET |
2 |
2 |
2 |
- |
2 |
2 |
521 |
536 |
|
| JOE |
2 |
2 |
- |
- |
2 |
2 |
522 |
548 |
|
| Yakima Yellow |
2 |
2 |
- |
- |
2 |
2 |
530 |
549 |
|
| Hex |
1 |
1 |
1 |
- |
1 |
1 |
535 |
556 |
|
| Cy3 |
- |
- |
3 |
- |
3 |
- |
546 |
563 |
|
| TAMRA |
- |
- |
3 |
- |
3 |
3 |
564 |
579 |
|
| ROX |
3 |
- |
3 |
- |
3 |
- |
576 |
601 |
|
| Texas Red |
- |
- |
3 |
- |
3 |
- |
586 |
610 |
|
| LC®Red 610 |
- |
- |
3 |
- |
3 |
- |
590 |
610 |
|
| LC®Red 640 |
- |
- |
3 |
3 |
- |
- |
625 |
640 |
|
| Cy5 |
- |
- |
3 |
3 |
- |
- |
646 |
662 |
|
| Cy5.5 |
3 |
- |
3 |
3 |
- |
- |
683 |
705 |
|
| IRD 700 |
3 |
- |
3 |
3 |
- |
- |
685 |
705 |
|
| *Please note that absorption and emission wavelengths are influenced by the solvent (pH, salt conditions, etc.), oligo sequence and other factors. |