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. Author manuscript; available in PMC: 2017 Mar 1.
Published in final edited form as: Curr Protoc Mouse Biol. 2016 Mar 1;6(1):39–66. doi: 10.1002/9780470942390.mo150178

Figure 2.

Figure 2

Quality control of the sgRNA and Cas9 mRNA preparations. (A) sgRNA seen on a nondenaturing agarose gel, stained with GelRed (Biotium). Lanes are samples 1 to 6, with sample 5 having a lower yield. Lane 7 is 1 Kb plus DNA ladder (Cat. No. 10787-018, Invitrogen). (B) Cas9 mRNA seen on a nondenaturing agarose gel, stained with GelRed. Lane 1: 2.5 μL withdrawn from the 96 μL in vitro transcription reaction; lane 2: 2.5 μL withdrawn from the 100 μL after poly adenylation was completed; Lane 3, 2.5 μL withdrawn from the 50 μL elution volume after purification with the MEGAclear kit. Lane 4: blank; Lane 5: 1 Kb plus DNA ladder. (C) Cas9 mRNA examined by spectrophotometry. The A260/A280 ratio is 2.3 and A260/A230 ratio 2.45 for this sample. The concentration is 1,125 ng/μL.