The gene of interest is cloned into one of the ITR/MCS-containing AAV vectors to generate AAV-GOI. The purity and RNAse contaminants of AAV viral plasmid should be taken into consideration.
Design genomic DNA primers that are approximatley 18 to 22 basepairs in length and have 45-55% GC content. For best results, use primers with a Tm greater than 55 C. Design primers to yield amplicon length between 400-500 bp. In addition, design primers so that the predicted cleavage site is not in the center of the amplicon and the detection reaction will yield two distinct product bands.
Genemedi has launched a comprehensive AAV packaging service combined with CRISPR/Cas9, the versatile genome-editing platform. The followings are some protocols of CRISPR /Cas9 mediated gene knockout in vitro and in vivo.
The gene of interest is cloned into one of the ITR/MCS-containing adenovirus vectors to generate pAd-GOI. The purity and RNA contaminants of viral plasmid should be taken into consideration.
This protocol is for the stable cell line construction based on puromycin selection.
The gene of interest is cloned into one of the LTR/MCS-containing lentivirus vectors to generate pLV-GOI. The purity and RNA contaminants of viral plasmid should be taken into consideration.
Animal experiment is essential in biomedical research. It contains establishment of a nude mouse tumor model and other disease model. Besides, gene transfection in vivo is included as well.