Compartmentalization in a Water-in-Oil Emulsion Repressed the Spontaneous Amplification of RNA by Qβ Replicase

During RNA replication mediated by Q beta replicase, self-replicating RNAs (RQ RNAs) are amplified without the addition of template RNA. This undesired amplification makes the study of target. RNA replication difficult, especially for long RNA Such its genomic RNA of Q beta phage. This perhaps is one of the reasons why the precise rate of genomic RNA replication In the presence of host factor Hfq has not been reported in vitro, Here, we report I method to repress RQ RNA amplification by compartmentalization of the reaction using a water-in-oil emulsion but Maintaining the activity of Q beta replicase. This method allowed LIS to amplify the phage Q beta genome RNA exponentially Without detectable amplification of RQ RNA. Furthermore, we found that the rate constant of genome RNA replication in the exponential phase at the optimum Hfq concentration was approximately 4.6 times larger than that of a previous report, close to in vivo data. This result indicates that the replication rate in vivo is largely explained by the presence of Hfq. This easy method paves the way for the Study of genomic RNA replication Without Special Care for the Undesired RQ RNA amplification.