A 1590 bp long CYP2C9 PCR product was generated using forward primer 5’-AGAAACCGGAGCCCCTGCAT and reverse primer 5’-AGAAGGCCAGTTCATCTCTATGTGC. The resulting 1590 bp PCR product was sequenced in both directions with the 5’-AGAAACCGGAGCCCCTGCAT (forward) and 5’-AGTTATGCACTTCTCTCACCCG (reverse) primers and aligned to NG_008385.2 to confirm the presence of g.48211A>T (NM_000771.4:c.1147A>T; p.K383X). Sequencing was performed using BigDye chemistry and a 3730 XL-DNA analyzer instrument (Applied Biosystems, Foster City, CA).
3730 xl dna analyzer instrument
Manufactured by Thermo Fisher Scientific
The 3730 XL-DNA analyzer instrument is a capillary electrophoresis system used for the automated analysis of DNA samples. It is designed to perform high-throughput DNA sequencing and fragment analysis.
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Lab products found in correlation
2 protocols using 3730 xl dna analyzer instrument
1
CYP2C9 Genotype Confirmation
2
Aptamer Sequence Determination and Structure Analysis
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Based on the highest MFI, the best round was selected. Briefly, the ssDNA of the selected round was amplified with the unlabeled forward and reverse primers. TA-cloning method was used for PCR products cloning. Next, to confirm the insertion of aptamers, PCR was performed with M13 and T7 primers on extracted recombinant plasmids. Then, nucleotide sequencing was done on the recombinant plasmids with proper insertion (Macrogen Co., Seoul, South Korea). Plasmids were sequenced based on M13 and T7 primers by the Applied Biosystems 3730XL DNA Analyzer instrument. Aptamer sequences were analyzed with BioEdit alignment software (version 7.2.5). Finally, aptamer structures were mapped and predicted by web-based software (M-fold: www.unafold.rna.albany.edu ; QGRS Mapper: www.bioinformatics.ramapo.edu ).
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