Hepes sucrose in extract
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Background: As part of the initiative in the “Containers” group in Build-a-Cell (https://github.com/BuildACell/liposome-kit/blob/master/txtl-liposome_water-in-oil.md). Within the protocol of encapsulation it has been determine that the internal solution should be composed of 100 mM HEPES and 200 mM sucrose. Initially, we work to encapsulate water-soluble fluorescent dye (HTPS or calcein) but looking towards the future of encapsulation TXTL a simple experiment was run to test the effect of 100 mM HEPES and 200 mM sucrose on the expression of deGFP.
Experiment: Experiment was run with a total TXTL reaction of 10 uL, with three different extract (Shaobin: donated by Shaobin Guo-Murray Group, ZJ: Zoila Jurado-with and without dialysis of extract). The positive control plasmid (OR1OR2-BCD2-deGFP) was obtain from lab stock. The negative control did not have plasmid present.
Result: Illustrate a negative effect of 100 mM HEPES and 200 mM sucrose on the expression of deGFP in all extract tested. Curious if this negative effect is as a result of the HEPES, sucrose, or crowding. Though when using TXTL HEPES should be omitted, and this is just more reason why.
