Multiplex QF-PCR Kit for Rapid Diagnosis of Trisomy 21, 18, 13 and Sex Chromosomes Aneuploidies
Five-Dye DNA Fragment Analysis
The Aneufast™ QF-PCR Kit uses a five-dye fluorescent system for automated DNA fragment analysis. This allows multiplex amplification and electrophoresis of over fifteen loci simultaneously. The kit is intended for use on Applied Biosystems ABI PRISM® genetic analysis instrumentation. Fluorochromes include 6-FAM™, VIC™, NED™ and PET™ to be used in conjunction with GS 500 LIZ™ size standard (Applied Biosystems PNº 4322682)
Hot Start Taq Polymerase and optimised PCR buffer
In order to maximise specificity of Multiplex PCR, Hot Start Taq Polymerase is included in the optimised PCR reaction buffer. The enzyme is completely inactive at room temperature. This prevents mis-priming during PCR set up. Activation is obtained during the 15 min. at 95ºC step before PCR cycling. This simplifies PCR set up and handling that can easily be done at room temperature.
Fluorescent primers should be stored away from light. The Aneufast™ box is internally coated with aluminium in order to achieve maximum light protection.
Aneufast™ is stable for up to one year if stored at -20ºC.
The Aneufast™ QF-PCR Kit
The two Multiplexes QF-PCR Sets S1 and S2 allow simultaneous analysis of four STRs on each of the autosomes 21, 18 and 13 in addition to sexing, using two pseudoautosomal STRs together with Amelogenin (AMXY) and SRY. These non -polymorphic sequences for sexing (AMXY and SRY) are independently amplified. Following collection of the products and simultaneous electrophoretic analysis, results from the S1 and S2 marker kits should be in agreement.
MXY, M21, M18 and M13
Chromosome- specific back-up marker sets are also available. MXY contains five STRs and two sexing markers markers for sex determination. M21, M13 and M18 contain five STRs on each of chromosomes 21 and 18, and four markers on chromosome 13. The back-up sets may be used either independently or in cases where all the S1 and S2 markers on any one of these chromosomes have been found to be uninformative (homozygous).
Note that in each chromosome- specific Set two markers amplified in S1 and S2 are repeated. This provides an opportunity to confirm sample identity. The chromosome- specific back-up sets may also be used to confirm any abnormal results. The inclusion of more markers increases the accuracy of the diagnosis.
For a comprehensive overview of the Aneufast™ kit our download page contains the full instruction manual and trouble-shooting guide.
Copyright Aneufast™ QF-PCR 2006
Most markers included in Aneufast™ have been extensively validated and applied on over 25.000 clinical specimens.