lentiCRISPR v2

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	There is no need to perform a negative control golden-gate reaction (without insert) as it will always contain colonies so not a good indicator of cloning success.

Protocol tips
There is no need to perform a negative control golden-gate reaction (without insert) as it will always contain colonies so not a good indicator of cloning success.

Publication protocol

Plasmids, antibodies and cell lines
pLentiCRISPR.v2, pAdeasy-1, pShuttle-CMV and pLentiLox3.7 (pLL3.7) were from Addgene. pNL4-3 BaL + (R5-tropic, containing the backbone of NL4-3 and the Env of BaL, referred to as BaL hereafter) and pNL4-3 (X4-tropic), as well as TZM-bl and C8166-CCR5 cell lines were from NIH AIDS Research & Reference Reagent Program, Division of AIDS, NIH. pCH042, a transmitted/founder HIV-1 molecular clone were kindly provided by Professor Beatrice H. Hahn ( Parrish et al., 2013 ). Anti-CCR5-PE (clone 2D7) and matched isotype control antibodies were from BD Pharmingen. Rabbit anti-Cas9 polyclonal antibody was purchased from Hangzhou HuaAn Biotechnology, and the internal control antibody (anti-β-actin) was purchased from Santa Cruz. CHO-CCR5, a CHO-K1 cell line with stable CCR5 expression was described previously ( Hu et al., 2005 ).

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Reviews

Discussion

Papers

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Manufacturer protocol

Download the product protocol from Addgene for lentiCRISPR v2 below.

Download manufacturer protocol

Videos

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