We first encapsulate hydrophilic reagents, such as calcein, rhodamine B and siRN

Question

We first encapsulate hydrophilic reagents, such as calcein, rhodamine B and siRNA, into the water core of RNV by microfluidic chips, and characterize the entrapment efficiency and the size of RNV. The entrapment of calcein (or rhodamine B) into the RNV and the entrapment efficiency are investigated using the spectrofluorophotometer and the microplate reader (Figure 1 B,C and Figures S2 and S3). To validate the encapsulation of siRNA into the RNV and quantify the entrapment efficiency of siRNA, we compare the gel retardation assays of free siRNA, lysed RNV loaded with siRNA (RNV is lysed by three cycles of freeze–thaw and sonication), and untreated RNV loaded with siRNA (RNV- [siRNA], Supporting Information). The amount of siRNA released from the lysed RNV[siRNA] (the middle band) is almost the same as that of free siRNA control (the left band), while free siRNA signal (the right band) in the untreated RNV

[siRNA] solution is barely observed, indicating the high siRNA recovery rate ( 98%) and loading efficiency in RNV (Figure 1D). The mea

I WANT TO ASK WHY ???THE RIGHT LINE IS UNTREATED rnv???

Explanation / Answer

The first band from the left is only siRNA.

Middle band is lysed RNV with siRNA. Hence it also gives a band similar to the first once, because RNV is lysed here, which leads to no encapsulation of siRNA.

But the third band from left has very low grey value, which is because here the siRNA is encapsulated into the RNV. RNV here was untreated which means it was intact alongwith the siRNA, giving low access to the siRNA on gel ultimately causing low grey value.

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