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Cas9protein,incomplexwithguideRNA,offersbenefitforCRISPR/Cas9technologyoverplasmidormRNAtransfection.ThebiggestbenefitisthatyoucanmakeRNPcomplexthatisreadytoworkinvivo.Cas9proteinisshortlived(about24hoursofhalflifeincells),thusminimizetheoff-targeteffectsandmosaicism.YoucanalsoeasilytesttheactivityofguideRNAinvitrobyasimplecleavageassay.Inaddition,Cas9proteinisprettystable,andactivityconfirmationismucheasiercomparedtomRNA.Lastly,itisactiveinmorebroadrangeoforganismssuchasnematodes,insects,plants,andnon-typicalmodelsystemsasitdoesnotrequiretranscriptionortranslation.
Cas9proteinofferedbyPNABioisconjugatedwithNLS,anditisinjection/transfectionreadyform.
Cy3-Cas9canbeaneasyenrichmentofthecellsthataretransfectedwithCRISPR/Cas9complex.
Invitrocleavageassay
PCRproductwasincubatedwithvariousamountsofCas9proteinandrunonthegel.InvitrocleavageassayisasimplewaytochecktheactivityofsgRNA.
TransfectionofRNPcomplex
RNPcomplexofsgRNAandvariousamountofCas9proteinwastransfectedintoNIH3T3cellsbyelectropration(Neon)orbylipofectamine(LF2000).
InjectionofRNPcomplex
Formouseembryoinjection,RNPcomplexof60ng/ulofCas9proteinand15ng/uleachoftwosgRNAswasinjectedintomouseembryo.ThetailDNAwaspreppedfromthepupsandsubjecttoT7E1assay.GreyrepresentsWT,greenforlargedeletion(#2,3,4,8),redforsmallin/del(#5,6,7),blueforhomozygoussmallin/del(#9,10).
matriks/Shikari® (S-ATP) Anti-Pembrolizumab ELISA w/confirmation/PEM-QNS-KEY
intactgenomics/Methionine Auxotrophic LBA4404 | ig® | Intact Genomics/15x50μl/1076-15
goprolytix/Proteolytic Activation of Prothrombin/1 mg, 200 µg/BCT-DFP
novateinbio/Dynorphin A ( 13-17 ) Porcine Peptide/5mg/PE-54024_5mg
matriks/Shikari® (Q-ETA) Etanercept ELISA/ETA-FD-ENB
intactgenomics/Agrobacterium ElectroComponent Combo Pack | Intact Genomics/12x50µl/1290-24
Jackson/Peroxidase AffiniPure Goat Anti-Mouse IgG (H+L)/2.0 ml/115-035-003
Jackson/Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L)/2.0 ml/111-035-003
Qubit™ dsDNA BR Assay Kit Q32853现货
Invitrogen Q32855 Qubit™ RNA HS Assay Kit现货促销
LUCK-1G,D-Luciferin, Potassium Salt 钾盐现货
GoldBio/D-Luciferin, Sodium Salt (Proven and Published™)/LUCNA-1G/1 g