...the challenge of trying to figure out what system might work best for your particular purposes. We’re here...than others. In today’s post, we will focus on a high-level comparison of popular cloning methodologies...results. If you’re in a hurry, Golden Gate cloning might be for you. This methodology relies on versatile... in as little as 30 minutes! Alternatively, you might consider Gibson cloning. In this technique, you ...fragments in an order determined by their homology. You might also consider FastCloning, first described by Chaokun...approach should you use if the budget is particularly tight? Most cloning processes will require a PCR reaction... job done, provided that you’ve already got the right enzymes laying around the lab. If not, you’ll be...

...and wrap the column with blue LED lights. When you switch off the lights, OptoNBs bind YFP and non-specific... the blue lights. Which option do you choose?  Option #2 may sound like fiction, but light-controlled...dark-induced dark-induced Loop 6 blue-light induced dark-induced blue-light induced Table 1: Summary of the... effect of AsLOV2 insertion site on light-induced binding.  Light controlled protein purification with...while cycling between dark and blue light which revealed a blue light-dependent shift in mCherry fluorescent...cells: light-based control of signaling pathways. This is particularly exciting because using light to control...binding of their target protein is altered by blue light illumination. This ability to reversibly bind and...

...conducted in Professor Cepko's lab and to get some insight into her management and mentoring styles. If you'd...

...pre-assembled genetic circuits such as logic gates and higher level gene networks. Sensing and Signaling Browse...Efflux Pumps Voigt Lab Repressors , Terminators , Light Signaling , Sigma Factors , Resource Allocator and...

...their large packaging capacity (>8 kb), high titers, and high levels of transgene expression. Additionally...

...γ-retroviruses. γ-Retroviruses can package relatively high amounts of DNA (up to ∼8 kb), and infection results...pRXTN CMV/MSV pRXTN is a modified version of pRX-tight Puro encoding NAT instead of PAC Iglehart 20672 ...

...Method Full Target Site (5' to 3') Left half-site Right half-site CACNA1F OZ507 and OZ508 OPEN OPEN gCGTGACCTCgctgatAAGAAGAGg...engineering of customized zinc-finger nucleases for highly efficient gene modification. Maeder ML et al. Mol...

... gRNA vectors suitable for prime editing are highlighted in the table below. Use the search bar to find...

...consider when planning experiments). What resources might be useful to help improve this technology or its...are some current obstacles that researchers face right now (e.g., not enough protocols, cost, not enough...

... (blue light-inducible AraC dimers in E. coli) that turns expression on and off with blue light.   Inspired...domain with VVD, a light-responsive domain that dimerizes upon exposure to blue light (Fig. 1). Another...VenturaIf you follow Barbara Di Ventura on Twitter, you might have seen the video of her singing about the paper...Another blue light-inducible dimerization domain, Vaucheria frigida aureochrome1 (VfAu1), led to functional...most applications.   Figure 1: With blue light, the VVD domains of the BLADE dimerize and activates...under the pBAD promoter, can be controlled with light without the need of any cloning.     What advantages...BLADE activates transcription with the flick of a light switch and shuts off just as quickly. The original...

... focused on three different levels of assembly: High efficiency and accurate single-insert assembly Intermediate...destination plasmid. Antibiotic selection allowed for high throughput cloning efficiency measurements. We performed...confirmed the insertion of the Lambda insert at the same high frequencies as the antibiotic selection markers....lacZ cassette assembly was easily achievable with high levels of transformants and low backgrounds – so...Using these 4 bp overhang observations, accurate “high-fidelity” junction sets for both the 12- and 24-...complex assemblies were performed. It was found that high efficiencies and accurate assembly levels were indeed...in the number of transformants while maintaining high levels of correct assemblies) translate to faster...

... 87429 TJP1-mEGFP AICSDP-23 mEGFP Tight junction protein ZO-1 Tight junctions 91565 AAVS1-mEGFP AICSDP...

...Human Platelet Receptors You may also like... Gavin Wright Lab plasmids This is a collection plasmids for ...Platelet Receptor Protein Microarray Identifies the High Affinity Immunoglobulin E Receptor Subunit α (FcεR1α...

...This is where FLEx enters the scene. FLEx: A light when night is about you! The FLEx (for flip-excision)...optogenetics? Optogenetics: Controlling neurons with light In 2005, a team from Stanford University, led by...ago with the discovery of microbial rhodopsins, light-sensitive proteins that absorb photons for energy...maintain membrane potential in response to visible light (Zhang et al., 2011). What do microbial rhodopsins...2000). Most neurons in the brain are not naturally light-sensitive, so the selective expression of opsin ...Optogenetic neuromodulation is made possible by the light: when hit with the correct wavelength, microbial..., 2011; Mei and Zhang, 2012). For example, blue light activates channelrhodopsins that, in turn, trigger...

...and C terminus and induces mutation at rates much higher than the parental vectors. pC-GoldyTALEN directs...obligate heterodimers that require cloning of left and right TALEN monomer proteins into opposite vectors. 37276...

...both P-elements and attB site for high expression of UAS-driven transgenes Worm unc-54,...Gateway cloning vectors Adenoviral High transduction efficiency, but do NOT integrate into...

...showed that improved specificity of Cas14 enables high-fidelity SNP genotyping. The development of  Cas14...

...Scientist. I used to spend my days (and often my nights :D) engineering E. coli to produce biofuels in ... screaming from the lab like the cartoon on the right?!? I would love to tell such a dramatic story, but...This may come as somewhat of surprise to many of my high school friends. In my teenage years I was super ...can involve walking alongside it and giving it a high five. While screaming can be therapeutic and I’ve...

...Parkinson's Richard Youle 23966 pTreTight-Htt94Q-CFP HTT CFP TREtight Huntington's Nico Dantuma 23971 ...Alzheimer's Gavin Wright 51643 APP-bio-His APP His, CD4 CMV Alzheimer's Gavin Wright 51811 PRNP-bio-His...PRNP-bio-His PRNP His, CD4 CMV Dementia Gavin Wright 53363 pCB268 APP Flag, HA, Myc CMV Alzheimer's Alexander Varshavsky...132389 AAVS1 CAG rtTA3 TauWT 2N4R-EGFP MAPT GFP TREtight Parkinson's, FTD Gerold Schmitt-Ulms 132393 AAVS1...AAVS1 CAG rtTA3 TauP301L 2N4R-EGFP MAPT GFP TREtight Parkinson's, FTD Gerold Schmitt-Ulms 133320 pHBS834...

...engineering using the CRISPR-Cas9 system. Ran FA, Hsu PD, Wright J, Agarwala V, Scott DA, Zhang F. Nat Protoc . ... engineered CRISPR-Cas9 complex. Konermann S*, Brigham MD*, Trevino AE, Joung J, Abudayyeh OO, Barcena...