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...it all together The FRET donors and acceptors described above can be paired together to measure a wide...

...Fact! Even though the Cas9:RNA complex is well described in the closed conformation, recent electron microscopy...

...transcriptional activators p65 and VP64 which in turn transcribes N-Cas9. The N- and C-Cas9 fragments then dimerize...

...named ABEmax. In 2020, two papers were published describing additional ABEs evolved from ABE7.10 that have...

...hot plasmids segment on the Addgene Podcast. Subscribe to the Addgene blog to get all the latest hot ...

... Bacterial Culture Introduction This protocol describes methodology for plating antibiotic over-agar for...

...Protocols Viral Service Introduction This protocol describes a general method for transfecting mammalian cells...

...gene of interest in cell culture. This protocol describes how to transfect suspension HEK293 cells with ...

... several μg/μL. The protocol below is meant to describe the general procedure for purifying plasmid DNA...

...from Feng Zhang (Addgene plasmid #52962 ) and is described in Improved vectors and genome-wide libraries ...

...plasmid of interest. For this example, we will describe how to copy a cDNA from one vector into a new ...

...Purification Protocol Introduction This protocol describes how to determine the purity and concentration ...

... Biology Reference Introduction This protocol describes how to use droplet digital PCR (ddPCR) to titer...

... Biology Reference Introduction This protocol describes how to extract RNA from tissues or cells without...

...Guide Virus Blog Posts Introduction This protocol describes how to use droplet digital PCR (ddPCR) to titer...

...license or rights of access or use to the materials described in the literature. The distribution of materials...

... polymerases that all transcribe different genes: RNA polymerase I — transcribes genes encoding ribosomal...of the RNA polymerase to DNA. RNA polymerase transcribes DNA to mRNA, which is ultimately translated into...upstream (5’) of the sense or coding strand of the transcribed gene (Figure 1). The coding strand is the DNA...noncoding strand, as this is the strand that is transcribed by the RNA polymerase. Figure 1: Simplified promoter...can bind), and the structural genes that are transcribed together. Operon regulation can be either negative... in the opposite way. The operon is normally transcribed until a repressor/corepressor binds to the operator...translated into a protein. RNA polymerase II — transcribes messenger RNA (mRNA), which is the RNA responsible...

...validate your genetic edit. Figure 1: Flow chart describing the general framework of a CRISPR experiment.... mRNA and Proteins In addition to the methods described above, you can also directly deliver mature mRNA...cells. In this case, your target cells are not transcribing DNA to mRNA, nor do they need to translate mRNA...directly combine purified Cas protein and in vitro transcribed gRNA to form a Cas-gRNA complex. You then delivery...endosomal approach. Similar to the mRNA method described above, this method is also transient, as the RNPs...

... is described in more detail in the Multiplex Assays section below. The following table describes the ...of B cells isolated from animals immunized as described above are used to create hybridomas, which produce...primary antibody for your protein of interest, as described in the IP section above. Reverse the crosslink...

... made of RNA. This retroviral RNA is reverse transcribed into DNA before being integrated in the genome...encoded on RNA (Figure 1). This RNA is reverse transcribed into the provirus, the cDNA that will be integrated...lentiviral packaging methods . The three plasmids described above (envelope, packaging, and transfer) are ...