...the amount of DNA used in a ligation based on the length of the DNA to get a proper ratio of 3 available...found on the web. Just enter the concentration, lengths of your insert and vector, and what ratio you want...

...cells. The problem is that the only version of full-length cDNA you can find for YGOI is in a bacterial expression...in the same restriction enzyme buffer (see New England Biolabs for more information about restriction ...

... Glanville J, Wolf P, Frenzel A, Wong J, Koh XY, Eng H-Y, Lane D, Lefranc M-P, Clark M, Dübel S (2018)...molecular definition, and potential for further engineering as compared to polyclonal or monoclonal antibodies...

... Glanville J, Wolf P, Frenzel A, Wong J, Koh XY, Eng H-Y, Lane D, Lefranc M-P, Clark M, Dübel S (2018)...

...within tumours via focused ultrasound. Nat Biomed Eng. 2021 Nov;5(11):1336-1347. doi: 10.1038/s41551-021...antibody?  Similarities:  Generated against full length GFP and converted to recombinant form by the NeuroMab...these processes. The Liu and Petsko labs recently engineered hyperfolder YFP (hfYFP), a FP well-suited for...

...should generally have the following properties: Length of 18-24 bases 40-60% G/C content Start and end...

.... to ligate an annealed oligo insert of 50bp in length into a 5kb vector, mix 100ng of the vector with...

...commercially available restriction enzymes, see New England Biolabs' website . Last Upload: May 18, 2018 Protocol...

...cells for 13 days. lentiCas9-Blast was a gift from Feng Zhang (Addgene plasmid #52962 ) and is described...

...extension time can be adjusted according to the length of the target sequence. How do I design primers...

...loading control. 1 lentiCas9-Blast was a gift from Feng Zhang (Addgene plasmid #52962 ) and is described...

...until fully dissolved. *Pro-Tip* This step is challenging due to the high viscosity of PEG. Stirring under...

...temperatures, but you may need to increase your primer length and increase the Tm if you are trying to clone ...

... Glanville J, Wolf P, Frenzel A, Wong J, Koh XY, Eng H-Y, Lane D, Lefranc M-P, Clark M, Dübel S (2018)...empower the scientific community with the tools to engineer them. Given the ease with which scientists can... Gong B, Nguyen K, van der List D, Murray KD, Engebrecht J, Trimmer JS (2019) A toolbox of IgG subclass-switched...Monoclonal Antibodies for Clinical Use. In: Antibody Engineering. InTech Additional resources on the Addgene blog...

...Quality Control (QC) Pooled libraries can be challenging and expensive to ensure adequate quality, but...

...pluripotency and nuclear reprogramming. Adv Biochem Eng Biotechnol, 2009. 114: p. 185-99. PubMed PMID: 19343304...embryonic stem-cell lines from human blastocysts. N Engl J Med, 2004. 350(13): p. 1353-6. PubMed PMID: 14999088...

...their applications in neuroscience. Curr Opin Biomed Eng. 2019. 12:111-117. doi: 10.1016/j.cobme.2019.10.010... as bright as the other GEVI systems that use engineered FPs. Attempts have been made to optimize their...their fluorescence, which ultimately led to the engineering of the second type of these GEVIs: a FP-fused... voltage sensor. FP-rhodopsin GEVIs have been engineered to have kinetics comparable to its one component... transiently and for genomic integration. Lab-engineered viruses will package your genetic cargo in lieu...

...of mice via adeno-associated viruses. Nat Biomed Eng 4:97–110 . https://doi.org/10.1038/s41551-019-0501...gRNA together (~4.2 kb) into an AAV vector is challenging due to its packaging capacity of AAV (~4.7 kb... little room for additional regulatory elements. Feng Zhang's group previously packaged Cas9 and multiple...enough to package into AAV. At 984 amino acids in length, Cas9 from Campylobacter jejuni (CjCas9) is the...ADAR2DD(delta984-1090) are approximately 4.1 kb in length, allowing them to be packaged in AAV.   Split ...transgenes might seem daunting, it’s actually a challenge that the field has overcome with split AAVs. In...When a cell is transduced by both AAVs, the full length gene and/or protein is reconstituted. One way to...

...enabling technologies in synthetic biology. J. Biol. Eng. 2013; 7, 13. PubMed PMID: 23663447. PubMed Central...fragments of the chromosome, which was technically challenging. Other homology based technologies We've described...Central PMCID: PMC3874208 Other Resources: New England Biolabs Gibson Assembly Overview Additional Resources...Protocols Gibson Assembly® is licensed to New England Biolabs by Synthetic Genomics, Inc. ...

...mechanobiology: seeing is believing." Annu. Rev. Biomed. Eng. 10 (2008): 1-38. PubMed PMID:18647110. Additional...spectral properties upon exposure to a specific wavelength of light. PA-FPs can be activated from low fluorescent...brighter fluorescent state by exposure to a specific wavelength of light. The switch or photoconversion usually...multiple times by activating with two distinct wavelengths of light (see Table 2). The best known reversible...photoactivation) (6). Dronpa is photoactivated by blue wavelengths of light, causing emission of green fluorescence...another group of reversible PA-FPs. These were engineered to be the first reversible, monomeric red fluorescent...dynamics (fission, fusion, etc). For example, by engineering PA-GFP tagged to a mitochondrial matrix protein...