I just generated the Gecko V2 human sgRNA plasmid library from addgene. When I ran the count_spacers.py as python ../ScreeningProtocolsmanuscript/countspacers.py -f ZXGecko2AS1R1001.fastq -i humangeckov2librarya09mar2015.csv, the statistics.txt looks like: Number of perfect guide matches: 0 Number of nonperfect guide ma...
I'm planning to use a custom-made CRISPR knockout library to target approximately 300 human genes. I am going to follow the Nature protocols 2017 of Joung et al. that I found really helpful ! My questions are: Considering the number of genes that will be targeted, is it better to use 6 or 4 sgRNAs per gene (similarly t...
Recently, I have amplified Mouse CRISPR Knockout Pooled Library (GeCKO v2) as described in protocol. Because I didn't mix them in any process, I acquired two amplified half libraries. After that, I qualified these two half libraries by NGS sequencing, and decided to screen the entire library in further studies. However, sin...
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