Jun 22, 2018

Public workspaceTransfection of Salpingoeca rosetta

DOI
dx.doi.org/10.17504/protocols.io.h8bb9sn
  • David S. Booth1,
  • Heather Middleton1,
  • and Nicole King1
  • 1HHMI/University of California, Berkeley
  • King Lab
  • Protist Research to Optimize Tools in Genetics (PROT-G)
David Booth
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DOI: dx.doi.org/10.17504/protocols.io.h8bb9sn
Collection Citation: David S. Booth, Heather Middleton, and Nicole King 2018. Transfection of Salpingoeca rosetta. protocols.io https://dx.doi.org/10.17504/protocols.io.h8bb9sn
License: This is an open access collection distributed under the terms of the Creative Commons Attribution License,  which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it's working
Created: May 31, 2017
Last Modified: June 22, 2018
Collection Integer ID: 6115
Abstract
This collection of protocols describes the preparation of reagents for transfecting DNA into the choanoflagellate Salpingoeca rosetta using a Nucleofector 4d Device and SF kit from Lonza. Plasmids that use S. rosetta promoters and regulatory elements to drive the expression of luciferase and fluorescent protein reporters are being deposited at Addgene. 
Acknowlegements: Arielle Woznica and Laura Wetzel for meticulous editing of protocol and clarifying steps in the procedure. The Gordon and Betty Moore Foundation for supporting this research.
Files
Protocol
Icon representing the file Protocol A: Preparation of endotoxin-free carrier DNA for transfection
Name
Protocol A: Preparation of endotoxin-free carrier DNA for transfection
Version 1
, University of California, San Francisco
David BoothUniversity of California, San Francisco
Protocol
Icon representing the file Protocol B: Preparation of ATP for transfection
Name
Protocol B: Preparation of ATP for transfection
Version 1
, University of California, San Francisco
David BoothUniversity of California, San Francisco
Protocol
Icon representing the file Protocol C: Preparation of heparin for transfection
Name
Protocol C: Preparation of heparin for transfection
Version 1
, University of California, San Francisco
David BoothUniversity of California, San Francisco
Protocol
Icon representing the file Protocol D: Preparation of BSA for quenching pretreatment
Name
Protocol D: Preparation of BSA for quenching pretreatment
Version 1
, University of California, San Francisco
David BoothUniversity of California, San Francisco
Protocol
Icon representing the file Protocol E: Preparation of pretreatment buffer
Name
Protocol E: Preparation of pretreatment buffer
Version 1
, University of California, San Francisco
David BoothUniversity of California, San Francisco
Protocol
Icon representing the file Protocol F: Preparation of recovery buffer
Name
Protocol F: Preparation of recovery buffer
Version 1
, University of California, San Francisco
David BoothUniversity of California, San Francisco
Protocol
Icon representing the file Protocol G: Preparation of reporter DNA
Name
Protocol G: Preparation of reporter DNA
Version 1
, University of California, San Francisco
David BoothUniversity of California, San Francisco
Protocol
Icon representing the file Protocol I: Transfection of Salpingoeca rosetta with Lonza 4d nucleofector
Name
Protocol I: Transfection of Salpingoeca rosetta with Lonza 4d nucleofector
Version 2
, University of California, San Francisco
David BoothUniversity of California, San Francisco