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View additional product information for Oncomine™ BRCA Research Assay, Chef-Ready Library Preparation - FAQs (A32841)
20 product FAQs found
The number of samples that can be multiplexed on the Ion 530 Chip will vary depending on whether automated or manual library preparation is used. If using automated library preparation, 32 somatic samples or 32 germline samples can be multiplexed on the Ion 530 Chip. If using manual library preparation, up to 96 germline samples can be multiplexed on the Ion 530 Chip. This is because of the available Ion Xpress Barcodes.
8 somatic samples or 32 germline samples can be multiplexed on the Ion 318 Chip.
The Ion OneTouch 2 and Ion PGM instruments are compatible with Oncomine BRCA Research Assay, Manual Library Preparation. The Ion Chef/Ion PGM and Ion Chef/Ion S5 or Ion S5 XL instruments are compatible with Oncomine BRCA Research Assay, Chef-Ready Library Preparation.
The Oncomine BRCA Research Assay Chef Ready Kit contains the Panel, library reagents and the Ion Code barcodes.
32 reactions can be performed with the Oncomine BRCA Research Assay Chef-Ready Library Preparation.
If you are not using the Ion Library Equalizer Kit for library normalization, one of the following can be used (but the library should be at 100 pM):
- Ion Library TaqMan Quantitation Kit
- Qubit 3.0 Fluorometer or Qubit 2.0 Fluorometer
- Qubit dsDNA HS Assay Kit
- Agilent 2100 Bioanalyzer Instrument
- Agilent High Sensitivity DNA Kit
Find additional tips, troubleshooting help, and resources within our Next-Generation Sequencing Support Center.
We recommend the following DNA isolation kits:
- RecoverAll Total Nucleic Acid Isolation Kit for FFPE (Cat. No. AM1975)
- Ion Ampliseq Direct FFPE DNA Kit (Cat. Nos. A31133, A31136)
- MagMAX FFPE DNA/RNA Ultra Kit (Cat. No. A31881)
- PureLink Genomic DNA Mini Kit (Cat. Nos, K182000, K182001, K182002)
We recommend two methods for gDNA quantification:
- Qubit 3.0 Fluorometer (Cat. No. Q33216) with Qubit dsDNA HS Assay Kit (Cat. Nos. Q32851, Q32854)
- qPCR using TaqMan RNase P Detection Reagents Kit (Cat. No. 4316831)
Yes, we offer the Oncomine BRCA Research Assay, Chef-Ready Library Preparation for that purpose.
The sample can be FFPE DNA or gDNA.
The expected amplicon size with the Oncomine BRCA Research Assay averages 175 bp.
The Oncomine BRCA Research Assay covers all the coding regions in BRCA 1 and BRCA 2 except the UTRs.
Yes, the Oncomine BRCA Research Assay is compatible with FFPE samples.
There are 2 pools with a total of 265 amplicons; pool 1 has 133 amplicons, pool 2 has 132 amplicons.
Yes. The Oncomine BRCA Research Assay has been validated for germline mutation detection as well as somatic variant detection.
The Oncomine BRCA Research Assay can potentially be used on other Ion instruments but customers would be required to perform their own validation and verification experiments.
We have validated the Oncomine BRCA Research Assay for use on the Ion PGM and Ion S5 platforms. With the Ion PGM instrument, both manual library preparation and Ion Chef library preparation are supported, with Ion Chef templating and OT2 templating. With the Ion S5 instrument, both manual library preparation and Ion Chef library preparation are supported with Ion Chef templating (OT2 templating has not been validated).
The Oncomine BRCA Research Assay, Chef-Ready Library Preparation contains pools 1 and 2 at 2X concentration, pre-measured in barcoded Primer Pool tubes ready to load into an Ion AmpliSeq Chef Reagents DL8 cartridge.
10 ng of DNA per target amplification can be used with the Oncomine BRCA Research Assay. The DNA can come from multiple sources, including FFPE samples and whole blood.
Both the Oncomine BRCA Research Assay and the AmpliSeq ‘Community Panel' Oncomine BRCA Research Assay will be available. The Oncomine BRCA Research Assay is designed for use on FFPE samples and uses a 2-pool approach rather than 3-pool approach. Customers using blood as a starting material should see the same results using the new Oncomine assay vs. the Community Panel.