... Some researchers have used fluorescently tagged DNA-binding proteins to label certain loci, but this ...depending on the hairpin combination bringing the total number of individual colors to 6. A gRNA with all...

...state machines (RSMs), which take on a particular DNA-sequence configuration (state) based on the identity...defined by the sequence within a prescribed region of DNA termed the “register”. Each chemical input to an ...result of their recombination is the excision of the DNA between the sites (Fig. 1B below). Alternatively,...result of their recombination is the inversion of the DNA between them (Fig. 1A below). In the absence of extra...interrogate state with qPCR. The re-arrangement of DNA segments in each state of a register enables the ...instructional text file. What's available at Addgene? In total, we constructed and tested eight two-input RSMs ...

...epigenetic analysis and designed using stretches of DNA that were transcriptionally active in retinal cell...from Jüttner et al., 2019 with permission. In total, this library contained 226 AAVs, each with a different...

... how proteins interact with one another (or with DNA) to perform diverse biochemical tasks in the cell...-IN” bioinformatics tool for guide RNA and donor DNA design, and implemented our optimised protocols into...to cells: a guide RNA, Cas9 protein, and a donor DNA that contains the tag sequence flanked by homology...CRISPR experiments (in collaboration with Integrated DNA Technologies, USA). These synthetic RNAs can also...csRNP complex formation, a single-stranded donor DNA (ssODN) that contains the tag-encoding sequence and... annealed cr:tracrRNA, and single-stranded donor DNA. The success of the epitope tagging experiment depends...tag-positive cells (tag ICC) as a percentage of total cells in the field (DAPI). Using the ICC assay as...

...distant cousins of well-known Cas9. Cpf1 cleaves DNA in a staggered pattern and requires only one RNA ...Kim et al. Kim et al. also compared the ratio of total off-target to on-target modification for AsCpf1 ...Our CRISPR Guide Pages Check Out MpAgo from the Doudna Lab Find Validated gRNAs ...

... the antibody has an antigen binding site for a total of 2 binding sites per antibody. C) IgG’s are big...23495938. Additional Resources on the Addgene Blog DIY DNA ladders from Penn State University Use Nanobodies...

...Cas9’s second HNH endonuclease domain. Cpf1 cleaves DNA in a staggered pattern and requires only one RNA ...Kim et al. Kim et al. also compared the ratio of total off-target to on-target modification for AsCpf1 ...

... viability controls such as DNA binding dyes and amine reactive dyes. DNA binding dyes, such as propidium...create a histogram of CD19-containing cells versus total cell number to determine the number of B cells in...membranes as is the case with dead or dying cells. DNA binding dyes are not compatible with protocols that...amine reactive dye, such as LIVE/DEAD stain. Like DNA binding dyes, amine reactive dyes can only pass through...

... biotin ligase from A. aeolicus, which lacks the DNA-binding domain (233 amino acids) found in the E. ...Central PMCID: PMC3699407. Milo, Ron. "What is the total number of protein molecules per cell volume? A call...

...plasmids are transformed in E. coli with the donor DNA for editing to take place. Then plasmids are cured.... coli W strain, and 1 Tatumella citrea, with a total of 70 sites edited. We optimized the strategy for...

...sure you hear it all the time, but Addgene is a totally invaluable resource. I really appreciate the work... plasmid request came on September 8, 2004, for pcDNA-p160MBP, while the first request for a kit came ...

...psi packaging signal. Viral components typically total <1.5kb, which is rarely more than 30% of the transfer... created for a variety of applications including cDNA expression, shRNA-mediated knockdown, Tet and Cre-regulated...

... cells and media are harvested and purified. In total, this process takes 4-7 days, not counting the time...are present in an AAV prep. AAV samples are first DNaseI digested to remove any residual AAV plasmid that... account for the dilution of the sample for the DNAseI digest. Use good sterile technique when handling...

...of recombinant DNA technology has allowed molecular biologists to fuse fragments of DNA together in a ...simultaneously incorporate multiple single-strand DNA (ssDNA) oligonucleotides (oligos), and thereby rapidly...sequence. Should one of these random stretches of DNA contain an enhancer, it will activate its own transcription...consists of 77,406 individual gRNAs targeting a total of 20,121 genes within the human genome (the identity...process alone involves much literature searching, DNA synthesis, and plasmid assembly. Luckily, researchers... platform for generating both simple and complex DNA constructs Have you ever wished you could design...Assembly method, which allows for easy assembly of DNA fragments by utilizing regions of sequence homology...

...system The Cas9 protein is composed of an n-terminal DNA recognition domain and a mostly c-terminal nuclease...c-terminal (Cas9(C)-2xNLS) fragments facilitates target DNA cleavage upon co-expression and spontaneous self-...the functional Cas9 molecule can cleave the target DNA. The inducible split cas9 approach can also be used...successful in regulating gene transcription using DNA-guided optogenetic tools; however, targeting the ...To overcome many of the limitations intrinsic to DNA-guided systems such as TAL effectors or Zinc Finger.... These memories would be recorded in the cell's DNA under the right conditions, and could initiate specified...cellular processes or be observed at a later time via DNA sequencing or other readout. Cellular memory is accomplished...

...example, AAV-PHP.eB is typically administered at a total dose between 1 x 1011 and 5 x 1011 vg per animal...produce in high titer, but higher doses (>1 x 1012 vg total in mice) may increase the chances of inducing an...-X1.1 showed the greatest fold change in RNA and DNA levels (~25-fold and ~4-fold respectively) relative...

...specific compartment. If your proteins are nuclear- or DNA-binding proteins, you may need to sonicate your lysates...these proteins. That being said, normalizing to total protein expression is preferable, and more accurate...