How are the 4 bases distinguished by the software?
The individual nucleotides are determined as light signals following a specific dispensation order of each individual nucleotide (see Chemistry)
What is the maximum size that can be sequenced?
150 base pair (depending on assay)
What limits the maximum size that can be sequenced?
The maximum read length comes down to the ability of the polymerase to keep fully incorporating nucleotides and the apyrase to degrade excess nucleotides. A reduction in both processes will lead to the sequence shifting out of phase.
How long does the Qseq take to sequence 100 known base pair?
There are 60 seconds between each dispensation, therefore, it would take 100 minutes, plus the 30 minutes required for template preparation (2 ¼ hours).
What sort of consumable is the sample placed into?
A 48 well disc
Does the binding of the amplicon to the streptavidin coated bead occur in the well where the sample is placed?
What is the range of biotinylated PCR amplicon recommended?
60 – 400 base pair
Do all of the steps involved in the pyrosequencing occur in the well?
Can you run multiple assays per run?
Can you run multiple applications (e.g. methylation analysis) per run?
How many primers can be automatically loaded per run?
There are four sequence primer injectors available per run, however, up to five sequence primers can be multiplexed per injector, giving a total of 20 sequence primers per run (assay dependent).
What is the function of the optical drop detection sensor?
The optical drop detection sensor monitors for anomalies during each injector dispensation. Any issues are flagged either during a run or analysis.
Where does the customer need to buy the chemistry from?
The kits are purchased from QIAGEN and the beads are purchased from GE Healthcare
What are the improvements over previous pyrosequencing instruments?
No sample preparation station, significantly reduced hands on time, fixed cartridges, smaller footprint, and automated primer loading
Is the Qseq made in Australia?
When the Qseq is purchased how many software licences are provided?
You may install the software on as many PC’s as required
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