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sgRNA production

MLM3613 plasmid (Joung lab) -

Sequences of oligonucleotides for sgRNA generation


Component Company Product number
Pfusion High Fidelity DNA Polymerase New England Biolabs M0530L
mMESSAGEmMACHINE T7 kit Life Technologies AM1344
Poly(A) tailing kit Life Technologies AM1350
MEGAscript T7 kit Life Technologies AM1333
RiboRuler High Range RNA ladder Thermo Scientific SM1821
RNeasy Mini kit Qiagen 74104
QIAquick PCR purification kit Qiagen 28104

RNA injection

Component Company Product number
Femtotips II Eppendorf 930000043
Halocarbon oil 700 Sigma H8898
Halocarbon oil 27 Sigma H8773

High resolution melt analysis

Component Company Product number
Proteinase K Life Technologies 25530-049
Hotshot Diamond PCR mastermix Clent Lifescience HS002
Skirted 96 well PCR plates Clent Lifescience 4ti-0961
LCGreen Plus+ melting dye BioFire Diagnostics BCHM-ASY-0005