Joseph Miano

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Questions related from Joseph Miano

Tamoxifen inducible KO in the RAT?

Has anyone had success with tamoxifen (or 4-hydroxytamoxifen) inducible Cre drivers in the rat? We heard of cases where rats were killed with high doses Tmx (>100 mg/kg). Even at 20 mg/kg we...

20 June 2023 1,663 0 View

What is the best way to manage Western blots?

Increasingly, more articles are being retracted because of duplicate or otherwise dubious Western blots. Consequently, journals are asking for original Western blots and authentication of...

24 February 2021 8,830 3 View

Has anyone been successful with NgAgo genome editing either in the mouse or in cultured cells?

The NgAgo nuclease uses a 5' phosphorylated single-strand DNA guide to create a double strand break in genomic DNA. It has been proposed as a superior genome editing system over CRISPR-Cas9....

14 August 2016 2,332 5 View

Has anyone compared BLESS with GUIDE-seq for unbiased detection of off targets following CRISPR-Cas9 genome editing?

The these assays utilize methods of "tagging" DSBs in the genome followed by PCR amplification of the library of sequences and NGS. They both seem rather temperamental but I wonder who (beyond...

09 July 2015 2,337 0 View

What conditions and anti-FLAG antibody are proven to work in tissue sections of mouse?

We recently made a FLAG-tagged mouse using 3 component CRISPR. We can do ChIP and Westerns very well. However, detecting the FLAG epitope in mouse sections (either frozen or paraffin) have been...

01 January 1970 6,548 4 View