For detection of nucleotides, the key factors demon- strated here are the pore diameter, the pulling speed, and spring constant of the AFM tip. Besides, several issues that were not taken into account in our simulation model might also affect the nucleotide detection ability in the real experi- mental situations (see more details in Appendix I in supple- mentary material16). For instance, the physisorption interactions between DNA and graphene were not considered explicitly in our model, although it is surprising to note that the peak force shows the same order as the interaction strength between nucleobases and graphene calculated using density functional theory.32 In real experiments, this interac- tion may enhance the force signal significantly and improve the discrimination ability of the current proposal.