Connector Inversion Probe (CIPer) technology is a powerful method for multiplex amplification, and has been validated here in genetic pathogen diagnostics, typing, and antibiotic resistance detection arenas as a proof-of-concept.
Microsatellite instability (MSI) was evaluated by multiplex amplification and sequencing.
Using multiplex amplification and microarray, we identified 883 (78%) mRNA species with signals significantly higher than backgrounds (p < 0.05, Welch's t-test) in T98G cells under various conditions.
It displays multiple advantages, such as rapid and precise allele determination, low DNA template requirement, multiplex amplification and fluorescence-based detection, digitized results and utilization of the abundant genomic element, At present, more than 60 countries worldwide have established forensic DNA databases based on STRs, and these databases continue to grow rapidly.
With our current protocol, it is possible to include two thousand or more gene transcripts in a single multiplex amplification group, and to analyze all human gene transcripts using several multiplexing amplification groups.
In conjunction with a newly developed high-throughput multiplex amplification system and highly simplified experimental procedures, the system can be used to analyze >1,000 mRNA species in a single assay.
We subsequently combined up to 12 loci in a single multiplex amplification reaction and developed, without extensive optimization (see Methods), an efficient 76 locus single-run INDEL genotyping panel in Atlantic salmon (Fig. 1; Additional file 2).
The first multiplex amplifications were carried out using equal concentration of locus specific forward and reverse primers (0.2 μM each) to screen eight individuals.
Multiplex amplifications were performed in mixtures containing 1.5 mM MgCl2, 0.2 mM dNTPs and 0.05 U of Taq recombinant polymerase per µl.
