Abi prism 3730 l genetic analyzer

Variant of the esxA gene from MDR-TB isolates was determined using PCR at the Institute of Tropical Disease, Surabaya, Indonesia. DNA isolation for the esxA examination was conducted using a QiAmp DNA Kit (QIAGEN). The PCR solution consisted of the KAPA2G Fast Ready Mix PCR Kit as PCR Master Mix, a pair of LIC-ESAT-6 primers (R: 5'-GAG GAG AAG CCC GGT TGC CCT TTC GCT ATT CTA CG-3' and F: 5'-GAC GAC GAC AAG ATG ACA GAG CAG CAG TGG AAT-3'), nuclease-free water, and the DNA template.
The PCR product was sent to 1 st Base using an ABI PRISM 3730 × l Genetic Analyzer. The sequence results were analyzed using GENETYX Ver. 10. The PCR product was sent to 1 st Base, Singapore for sequencing. Sequencing of the esxA gene was performed using an ABI PRISM 3730 × l Genetic Analyzer with 96 capillaries developed by Applied Biosystems and a Big Dye ® Terminator v3.1 Cycle Sequencing Kit.
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Peripheral white blood cells were harvested for genomic DNA extraction. Polymerase chain reaction (PCR) and amplicon fragment length analysis were used to analyze the HTT CAG trinucleotide repeat length. The CAG repeat regions were amplified by PCR with one primer of the fluorescently labeled primer pair. The forward and reverse sequences of the PCR primers were 5'-6FAM-atgaaggccttcgagtccctcaagtccttc-3' and 5'ggcggtggcggctgttgctgctgctgctgc-3', respectively. Fragment length analysis of the amplicons was conducted on an ABI Prism 3730 × l Genetic Analyzer (Applied Biosystems, Waltham, MA, USA) using the GeneScan 500 LIZ dye Size Standard (Applied Biosystems) and GeneMapper software (v3.7; Applied Biosystems). The size of the alleles was used to calculate the CAG repeat numbers.