February 25, 2016 at 12:26AM

Today I Learned: 1) It really can't hurt to ask. 2) A mark above or below a character to change its pronounciation (like the degree symbol in å or the "'" in á) is called a "diacritic". 3) Most standard systems for making guide RNA plasmids involve a linearized plasmid backbone with a promoter at one end (pointing off the end) and a tracrRNA sequence at the other end. To build the vector, you buy phosphorylated primers with your target sequence and just blunt-end ligate (or, probably, restriction digest/ligate, depending on the system) it onto the ends of the backbone, and voila! You have a plasmid that produces a hybrid guide/tracr RNA!