The procedure of phase construction between rs11836367 and the reporter SNP rs2160989 was performed as described previously with modification (29 (link)). Genomic DNA from wild-type MCF10A was amplified using HiFi HotStart DNA Polymerase (KAPA Biosystems, Wilmington, MA, USA). PCR products were ligated into the PCR-linearized pUC19 vector using the ClonExpress II One Step Cloning Kit (Vazyme, Nanjing, China) according to the manufacturer’s instructions. The phase between rs11836367 and rs2160989 was manually determined on the basis of the genotype of a linked heterozygous SNP in the overlapping region between fragments. Details about the PCR primer sequences used are listed in table S7.
Protocol detail
Determining Genetic Phase Between SNPs
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Corresponding Organization : Peking University Third Hospital
Other organizations : University of California, San Francisco, Peking University Cancer Hospital, UCSF Helen Diller Family Comprehensive Cancer Center
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Variable analysis
independent variables
- Rs11836367
- Rs2160989
- Phase between rs11836367 and rs2160989
- Wild-type MCF10A genomic DNA
- HiFi HotStart DNA Polymerase (KAPA Biosystems, Wilmington, MA, USA)
- PCR-linearized pUC19 vector
- ClonExpress II One Step Cloning Kit (Vazyme, Nanjing, China)
- Positive control: Not explicitly mentioned.
- Negative control: Not explicitly mentioned.
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