Adult Ciona robusta (intestinalis Type A) were collected from San Diego, CA (M-REP). Dechorionated embryos were obtained and electroporated as previously established (Christiaen et al., 2009a , b ). Sequences of in situ hybridization probe templates can be found in Supplemental Table 3 . Neurog perturbation and control plasmids were previously published (Stolfi et al., 2015 (link)). Probes were prepared either from published clones, synthetic DNA fragments (Twist Bioscience, San Francisco, CA, United States), or directly from RT-PCR amplicons (see Supplemental Table 3 for details). Probe synthesis and fluorescent, whole-mount in situ hybridization were carried out as previously described (Beh et al., 2007 (link); Ikuta and Saiga, 2007 (link)). dpERK staining was carried out as previously described (Stolfi et al., 2011 (link)), using 1:500 mouse monoclonal anti-dpERK antibody (Sigma #M9692), and tyramide signal amplification.
Synthetic dna fragments
Manufactured by Twist Bioscience
Sourced in United States
Synthetic DNA fragments are laboratory equipment used for the production and manipulation of artificial genetic sequences. These fragments serve as the fundamental building blocks for various applications in fields such as molecular biology, genetic engineering, and synthetic biology. The core function of synthetic DNA fragments is to provide a customizable source of genetic material that can be utilized for a wide range of scientific and research purposes.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using synthetic dna fragments
1
Whole-mount in situ hybridization in Ciona
2
Yeast Histone Expression Vectors
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Yeast histone expression vectors were derived from the HHT1-HHF1 URA3 CEN ARS plasmid pMS329 (gift of Mitchell Smith) and the HHT2-HHF2 TRP1 CEN preprint (which was not certified by peer review) is the author/funder. All rights reserved. No reuse allowed without permission.
The copyright holder for this this version posted October 28, 2021. ; https://doi.org/10.1101/2021.10.27.466091 doi: bioRxiv preprint 14 ARS plasmid pWZ414-F12 (obtained from Rolf Sternglanz) (Megee et al., 1990; (link)Zhang, 1998) . The HHT1(R49C) vector (pEL649) and the HHT2(R49C) (pEL629) vector were constructed by digesting pMS329 and pWZ414-F12 respectively with ClaI/AgeI and with BlpI/BamHI to remove a segment of their H3 genes followed by recombination with synthetic DNA fragments (Twist Bioscience) containing the R49C mutation by the Gibson assembly protocol (New England Biolabs). Similarly, the 2xV5-HHT1 (pEL656) and 2xV5-HHT1(R49C) (pEL650) plasmids were generated by digesting pMS329 and pEL649, respectively, with AgeI/PmeI followed by recombination with a synthetic fragment containing the N-terminal 2xV5 tag. The integrity of all plasmids was confirmed by Sanger sequencing (Genewiz).
The copyright holder for this this version posted October 28, 2021. ; https://doi.org/10.1101/2021.10.27.466091 doi: bioRxiv preprint 14 ARS plasmid pWZ414-F12 (obtained from Rolf Sternglanz) (Megee et al., 1990; (link)Zhang, 1998) . The HHT1(R49C) vector (pEL649) and the HHT2(R49C) (pEL629) vector were constructed by digesting pMS329 and pWZ414-F12 respectively with ClaI/AgeI and with BlpI/BamHI to remove a segment of their H3 genes followed by recombination with synthetic DNA fragments (Twist Bioscience) containing the R49C mutation by the Gibson assembly protocol (New England Biolabs). Similarly, the 2xV5-HHT1 (pEL656) and 2xV5-HHT1(R49C) (pEL650) plasmids were generated by digesting pMS329 and pEL649, respectively, with AgeI/PmeI followed by recombination with a synthetic fragment containing the N-terminal 2xV5 tag. The integrity of all plasmids was confirmed by Sanger sequencing (Genewiz).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!