INFINIUM Assay and Genotyping B Chip Procedure

These flashcards cover key concepts and procedures involved in the INFINIUM assay used for processing genotyping B chips.

What is the overall goal of the INFINIUM assay procedure?

To successfully process a genotyping B chip.

What is the first step in the INFINIUM assay procedure?

Amplifying, fragmenting, and isolating the DNA.

What is hybridization in the context of the INFINIUM assay?

The process where denatured DNA strands bind to oligonucleotides attached to beads on the array.

How is the oligonucleotide attached to the bead prepared for detection?

It is stained and selectively extended by one base.

What should you do one day prior to amplification?

Prepare the DNA by dispensing 200 ng of DNA per well into a deep well 96 sample plate.

Why is it necessary to label the plate with a barcode sticker?

To ensure proper identification of samples and avoid mix-ups.

What should be done with the samples after dispensing into the deep well plate?

Centrifuge the plate and leave samples overnight to evaporate liquid.

How much MA1 reagent should be added per well?

Twenty microliters.

What is the purpose of incubating the plate for at least thirty minutes after adding reagents?

To allow the reagents to react and properly prepare the samples for amplification.

What indicates that DNA samples are successfully precipitated after centrifugation?

A blue pellet at the bottom of the plate.

What temperature should the samples be denatured at before hybridization?

At a specific temperature, which is not provided but implies a heat block is used.

What must be prepared before the completion of the cooling period?

The hive chamber for the bead chips.

What is critical during the loading of the Bead Chips?

Extraordinary care must be taken to place the correct sample on the correct bead chip at the correct position.

For how long should the HYBE chamber be incubated in an oven?

For 16 to 24 hours.

What should be done after removing bead chips from the oven?

They should be incubated at room temperature for 25 minutes to cool down.

What happens to the bead chips after removing the cover seal?

They are submerged in PB1 without touching the beads.

What is the importance of the scanner output in the GENOME STUDIO software?

It generates cluster profiles indicating genotype information.

What does a successful bead chip display during scanning reveal?

Bright and distinct red and green laser light intensities corresponding to the labeled nucleotides.

What must be avoided when processing B chips?

Exposure to air for extended periods.