Nanodrop 1000 spectrophotometer

Fecal DNA was extracted from stabilized samples, according to the instructions provided by the manufacturer using a spin-column based kit (Qiagen Fast DNA Stool Kit, Qiagen, Germany). Briefly, homogenized suspensions were heated to 95 °C for 10 min, centrifuged and treated with Proteinase K. Then, samples were incubated with the buffer provided in the kit and pure ethanol. After vortexing, the preparations were transferred onto spin columns and centrifuged, followed by multiple washing steps on the column using the washing buffer from the isolation kit. Finally, DNA was eluated in 100 µL of a TE-based buffer. Nucleic acid concentration was measured using a Nanodrop 1000 spectrophotometer (Eppendorf, Hamburg, Germany).

Venous blood of 3 ml was drawn from 22 CML subjects. The RNA was extracted by RNA method. The quantification of total RNA was performed using the NanoDrop™ 1000 Spectrophotometer (Eppendorf, USA) [32 ]. The cDNA was synthesized using the cDNA synthesis kit provided by Thermo Scientific (cat# 1621) as per manufacturer instruction [33 (link), 34 (link)]. The cDNA was used as a template in the first round of the PCR while the amplified product of the first round was used as the template for the second round of the PCR. The sequences of the primer used in the nested PCR reaction were in round 1 B2A (F) 5′-TTCAGAAGCTTCTCCCTGACAT-3′ and ABL 4065 (R) 5′-CCTTCTCTAGCAGCTCATACACCTG-3′ and in round 2 BCR F4 (F) 5′-ACAGCATTCCGCTGACCATCAATA-3′ and U396 (R) 5′-GCCATAGGTAGCAATTTCCC-3 followed from Akram et al. [33 (link)].