Direct genotyping from whole blood using alkaline polyethylene glycol

• Published in: Analytical biochemistry Vol. 582; p. 113351
• Main Authors: Liu, Xiaonan, Zhang, Chao, Hua, Kai, Liang, Jianping, Li, Hang, Ma, Ting, Zhu, Juanli, Cui, Yali
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.10.2019
• Subjects: Alkaline polyethylene glycol; Direct genotyping; Whole blood; Whole blood; Alkaline polyethylene glycol; PEG; Direct genotyping; MTHFR; GMNP; C line; RMU; T line; WT; M; APEG
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/j.ab.2019.113351

In order to simplify biological sample preparation to meet the demand of rapid genotyping, we improve alkaline polyethylene glycol (APEG) based on Chomczynski's procedure for the PCR-based lateral flow genotyping system, which enables the rapid and efficient direct genotyping from whole blood without DNA purification. The improved APEG has a high tolerance for extreme storage conditions. Testing whole blood with an abnormal hematological index indicates that APEG efficiency would not be influenced by pathological factors. Compared with sequencing, the accuracy of this genotyping system was 100% on testing with 200 clinical samples. [Display omitted] •Whole blood treatment employing alkaline polyethylene glycol for direct genotyping.•Simplify whole blood preparation for efficient release of genomic DNA from leukocytes.•Inhibit the effect of DNA polymerase inhibitors from blood.•Ignore whole blood with abnormal hematological indexs.•Stable leukocyte lysis buffer in long term storage.