The therascreen IDH1/2 RGQ PCR Kit provides reagents to perform 9 separate amplification reactions for detection of 12 mutations:
- 3 total amplification reactions of codons 132 and 100 of the IDH1 gene and of codon 172 of the IDH2 gene
- 3 mutation amplification reactions of codons 132 and 100 of the IDH1 gene and of codon 172 of the IDH2 gene
- 3 mutation-specific amplification reactions of IDH1 R132H, IDH1 R132C, and IDH2 R172K mutations
IDH1 and IDH2 mutations detected using the therascreen IDH1/2 RGQ PCR Kit
|Gene || Mutation || Base change ||Cosmic ID |
| Arg132His (R132H || 395G>A ||COSM28746 |
| Arg132Cys (R132C) || 394C>T || COSM28747 |
| Arg132Ser (R132S) || 394C>A || COSM28748 |
| Arg132Gly (R132G) || 394 C>G || COSM28749 |
| Arg132Leu (R132L) || 395G>T || COSM28750 |
| Arg132Val (R132V) || 394_395 CG>GT || COSM28751 |
| Arg100Gln (R100Q) || 299G>A || COSM88208 |
| Arg172Lys (R172K) || 515G>A || COSM33733 |
| Arg172Met (R172M) || 515G>T || COSM33732 |
| Arg172Trp (R172W) || 514A>T || COSM34039 |
| Arg172Ser (R172S) || 516G>T || COSM34090 |
| Arg172Gly (R172G) || 514A>G ||COSM33731 |
Total reaction mixes
The Total Primers and Probe Mixes (PPM-Total) use primers and probes to amplify both mutated and wild-type target sequences.
Mutation detection reaction mixes
The mutation detection primers and probe mixes combine primers and probes, to amplify both mutated and wild-type target sequences, plus an oligonucleotide, 3' blocked with the addition of a phosphate group to prevent elongation (PCR clamping), which is specific to the wild-type target sequence.
When the PCR template contains the wild-type sequence, the 3'-phosphate oligonucleotide will dominate over PCR primer binding due to higher affinity. There is no or low extension by the DNA polymerase and no or low amplification is observed.
When a mutated sequence is present, PCR primer binding will dominate over the 3'-phosphate oligonucleotide binding and amplification will proceed.
Mutation identification reaction mixes
Allele-specific amplification is achieved by ARMS (Amplification Refractory Mutation System), which exploits the ability of the DNA polymerase to distinguish between a match and a mismatch at the 3' end of a PCR primer.
When the PCR primer is fully matched, the amplification proceeds with full efficiency. When the 3' base is mismatched, only low-level background amplification occurs.