Calibration methods, equipment and computer program products
A technology of calibrator and calibration curve, which is applied in biochemical equipment and methods, sequence analysis, biostatistics, etc., and can solve problems such as consumption
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Embodiment 1
[0091] Example 1 describes how to establish fixed points for subsequent use in a single-point calibration procedure using a linear reference calibration curve generated by a first instrument that amplifies nucleic acid and monitors amplicon production in a real-time format. The first instrument is hereinafter referred to as instrument "V35".
[0092] Example 1
[0093] Establishes fixed points for internal calibration adjustments
[0094] The 6 nucleic acid calibration standards were amplified in 5 replicates using the first real-time instrument identified as V35. All reactions on Instrument V35 were performed using standard target capture reagents ("TCR") to enrich the target nucleic acid prior to amplification. The samples used for the calibration reaction have a volume of 0.5 ml each and a range of 10 2 to 10 7 copies / ml of nucleic acid target concentration. Nucleic acids captured from different samples were repeatedly co-amplified with fixed 150,000 copies of the IC...
Embodiment 2
[0102] Example 2 illustrates how results obtained on a second real-time instrument (ie hereinafter "V53") can be processed using fixed points established on a first real-time instrument (ie V35). The results obtained using Instrument V53 for the amplification of the different calibration standards were individually paired with established fixed points using Instrument V35 to create a linear calibration curve plot. These calibration graphs were then used to back check the results from the remaining reactions performed on the instrument V53. In other words, treat one calibration standard as the adjusted calibrator and the other calibration standard as the mock test sample, and use the adjusted calibration curve determined using the results from the adjusted calibrator and fixed points to determine the target present in the mock test sample. amount of nucleic acid.
[0103] Example 2
[0104] Plot an adjusted calibration curve incorporating fixed points
[0105] A nucleic ac...
Embodiment 3
[0116] Example 3 exemplifies how fixed points established by the methods described above can also be used in procedures for quantifying nucleic acid targets using reagents that are impaired by accelerated degradation.
[0117] Example 3
[0118] Quantify target nucleic acids using reagents subjected to accelerated degradation conditions
[0119] Six nucleic acid calibration standards were amplified using a third real-time instrument identified as V47. All reactions were performed using TCRs that had been heated at 55°C for 28 days to facilitate accelerated degradation. All other amplification reactions and monitoring conditions were similar to those described above.
[0120] Table 4 presents the summary results and value-assigned concentrations of the calibration standards, where all 6 calibrators were used for value assignment. The numerical results in Table 4 are presented using more than the appropriate two decimal places.
[0121] Table 4
[0122] Summary results of...
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