Does anyone help me out to clone the Oligo sequence in Lenti CRISPRv2 backbone plasmid ?

19 May 2017 1 7K Report

Hi.. Everyone I have been attempted to Clone the Oligo sequence into the LentiCRISPRv2 backbone plasmid but I'm unable to get the colonies when I transformed in DH5alpha. I have followed the Addgene protocol (http://genome-engineering. org/gecko/wp-content/uploads/ 2013/12/lentiCRISPRv2-and- lentiGuide-oligo-cloning- protocol.pdf).

Even I tried different time interval and ratio of vector: insert.

Thank you

Aamal Ghazi Mahdi Al-Saadi

You may also take a look to

http://www.origene.com/assets/Documents/Lentiviral/how_to_use_lentivirus_a_short_protocol.pdf

https://media.addgene.org/cms/files/Zhang_lab_LentiCRISPR_library_protocol.pdf

Good luck

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