<aside> <img src="/icons/light-bulb_gray.svg" alt="/icons/light-bulb_gray.svg" width="40px" /> Getting Started

</aside>

Go through this page before entering the lab to guide the design of a prototypical PURE inner solution to be used in liposomes.


<aside> <img src="/icons/merge_gray.svg" alt="/icons/merge_gray.svg" width="40px" /> Prerequisite Protocols

</aside>

Outer solution for PURE reactions

<aside> <img src="/icons/iterate_gray.svg" alt="/icons/iterate_gray.svg" width="40px" /> Protocol

</aside>


Inner solution for PURE reactions

Component Reaction Volume (ul) Total Volume (ul)
Master Mix (5x reactions)
NEB-Sol A 4 20
NEB-Sol B 3 15
RNAse Inb. 0.5 2.5
Sucrose (2 M) 1.5 7.5
Total 9 45

When specifying specific experimental conditions we use the following notation OS-T1T2...-IS, where OS = outer solution; T1, T2,… = DNA template 1, DNA template 2, …; and IS = inner solution. Here is an example where the DNA templates used contain sequences encoding for GFP and aHly:

OS-T1T2-IS OS-T2-IS notes
Master Mix 9 9
DNA-aHly 0.5 0.5 10 fmol/uL
DNA-eGFP 0.0 0.5 10 fmol/uL
nuc Free H20 0.5 0.0
Total 10 10

<aside> <img src="/icons/checklist_gray.svg" alt="/icons/checklist_gray.svg" width="40px" /> In progress

</aside>