IDH1/2 RGQ PCR Kit

For the detection of 12 IDH1 and IDH2 mutations in DNA from FFPE brain tumor tissue

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IDH1/2 RGQ PCR Kit (20)

Cat. No. / ID: 873001

For 20 reactions: 9 Primer and Probe Mixes, WT Control, Positive Control, Master Mix, Nuclease-Free Water
For Research Use Only. Not for use in diagnostics procedures. No claim or representation is intended to provide information for the diagnosis, prevention, or treatment of a disease.

✓ 24/7 automatic processing of online orders

✓ Knowledgeable and professional Product & Technical Support

✓ Fast and reliable (re)-ordering

Features

  • Accurate detection of 7 IDH1 and 5 IDH2 mutations using PCR clamping
  • Sensitive identification of 3 mutations using ARMS PCR technology
  • A standardized assay with ready-to-use solutions
  • An easy workflow with the Rotor-Gene Q instrument

Product Details

The IDH1/2 RGQ PCR Kit enables detection of 12 mutations in the IDH1 and IDH2 genes using real-time PCR on the Rotor-Gene Q 5plex HRM Instrument. The kit provides all necessary reagents, optimized for rapid and sensitive detection of IDH1 and IDH2 mutations.

Principle

The 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 IDH1/2 RGQ PCR Kit
Gene Mutation Base change Cosmic ID

IDH1
Arg132His 395G>A COSM28746
Arg132Cys 394C>T COSM28747
Arg132Ser 394C>A COSM28748
Arg132Gly 394 C>G COSM28749
Arg132Leu 395G>T COSM28750
Arg132Val 394_395 CG>GT COSM28751
Arg100Gln 299G>A COSM88208

IDH2
Arg172Lys 515G>A COSM33733
Arg172Met 515G>T COSM33732
Arg172Trp 514A>T COSM34039
Arg172Ser 516G>T COSM34090
Arg172Gly 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.

Procedure

The IDH1/2 RGQ PCR Kit provides all necessary reagents for detection of 12 mutations in the IDH1 and IDH2 genes. Nine primer and probe mixes are used to amplify both mutated and wild-type target sequences. Simply add template DNA and the appropriate primer and probe mix to the ready-to-use PCR master mix, and start the reaction on the Rotor-Gene Q 5plex HRM instrument using the optimized cycling program described in the kit handbook. For DNA purification, we recommend using the QIAsymphony DNA Mini Kit on the QIAsymphony SP or the QIAamp DNA FFPE Tissue Kit, according to the modified protocols in the IDH1/2 RGQ PCR Kit Handbook.

Applications

The IDH1/2 RGQ PCR Kit enables detection of 7 mutations in the IDH1 gene, 5 mutations in the IDH2 gene, and identifies 3 major mutations, in DNA extracted from formalin-fixed, paraffin-embedded (FFPE) human brain tumor tissue specimens.

Resources

Safety Data Sheets (1)
Download Safety Data Sheets for QIAGEN product components.
Kit Handbooks (1)

FAQ

Can I use stained slide materials with IDH 1/2 RGQ PCR kit?

No.  The pathologist should only proceed with unstained material.

 

 

FAQ ID -3101
Can I perform the IDH 1/2 RGQ PCR test with slides containing less than 40% tumor cells?

No. To detect all mutations, it is required to have at least 40% tumor cells.

FAQ ID -3098
Can I use the QIAGEN deparaffinization solution with the IDH 1/2 RGQ PCR kit?
No. The QIAGEN deparaffinization solution does not work with our kit.
FAQ ID -3100