mMessage mMachine Help

Hi all,

Hoping to get a bit of help with the SP6 mMessage mMachine kit. I’ve been making mRNAs with this kit for micro injections, and it’s been going fine, I get expression off of the transcripts I inject, but I’m unhappy with the quality of the mRNA I produce. Some transcripts give perfect clean slim peaks by tapestation, but others I get real broad peaks. Same data but by gel, I’ll get some with clean bands with just a dusting of smear below it, but some band is barely visible with just a smear. Same result when I use the kit control template. Both express fine, but I’d love to be injecting as a pure a product as possible. I’ve worked with rna a lot, I’m RNA zapping everything in site, nuclease free water, all the good stuff, zymo clean up kit after the reaction. Any suggestions on how to make higher purity transcripts? I do run the rxn at 30c O.N., 37 for 1hr per kit sop was way worse. Transcribing off of gel purified (qiagen) cut plasmid.

Thank you so much!