BioMatrix with 192 plate capacity rotating carousels.
Yiqun Chen re-loading a BioMatrix.
A BioMatrix replicating 1536 yeast colonies per plate.
Automated Plate Pourer by KREO technologies
Protocols
We have made a number of improvements on SGA strains and methods and the following sections describe the latest improvements.
SGA Methodology
We have manuscripts describing SGA methodology:
- A. Tong and C. Boone (2005)
Synthetic Genetic Array (SGA) Analysis in Saccharomyces cerevisiae.
Yeast Protocols, Second Edition. Methods in Molecular Biology, 313:171- 92. (The Humana Press Inc., Totowa, NJ, U. S. A . )
Download PDF document. - A. Tong and C. Boone (2007)
High-Throughput Strain Construction and Systematic Synthetic Lethal Screening in Saccharomyces cerevisiae.
Yeast Gene Analysis - Second Edition. Methods in Microbiology, Volume 36: 369-386, 706-707 (Elsevier Ltd.)
Download PDF document. - Freezing down 96 well format libraries.
Download XLS document. - Boone SGA array definition files
- Standard SGA array (in 384 density format)
Download XLS document. - Slow Growing array (in 96 density format)
Download XLS document.
SGA Strain Kit
Upon request, we send out an SGA kit that contains: p4339, for switching kanMX deletion alleles to natMX alleles; Y7092, our standard SGA starting strain, into which query mutations are introduced by transformation; Y8205, another SGA starting strain, into which query mutations can be introduced by the switching protocol.
p4339
TA::MX4-natR switcher cassette; This plasmid can be used to switch from kanR KO kan-MX or URA3 KO URA-MX to natR KO. Cut with Eco RI to target.
P4339 was constructed by amplifying natMX [A. L. Goldstein, J. H. McCusker, Yeast 15, 1541 (1999)] with primers (5'-ACATGGAGGCCCAGAATACCC-3' and 5'-CAGTATAGCGACCAGCATTCAC-3') and the product was introduced into pCRII-TOPO (InVitrogen) at the unique EcoRI sites.
E. coli strain: TOP10F
Plasmid is ampicillin resistant and should be cultured in 2YT + 120ug/ml ampicillin.
Y7092
Starting strain for SGA query strain construction by PCR knockout.
Mating type: MAT alpha
Genotype: can1delta::STE2pr-Sp_his5 lyp1delta his3delta1 leu2delta0 ura3delta0 met15delta0
(Note: "STE2pr " represents the MATa-specific STE2 promoter; "Sp_his5 " represents the S. pombe his5 , which complements S. cerevisiae HIS3 ; also, this strain carries a wild-type LYS2 )
Other SGA starting strains carrying different reporters are available upon request.
Y8205
Starting strain for SGA query strain construction by switching.
Mating type: MAT alpha
Genotype: can1delta ::STE2pr-Sp_his5 lyp1delta ::STE3pr-LEU2 his3delta1 leu2delta0 ura3delta0
Upon request, we send out an SGA kit that contains: p4339, for switching kanMX deletion alleles to natMX alleles; Y7092, our standard SGA starting strain, into which query mutations are introduced by transformation; Y8205, another SGA starting strain, into which query mutations can be introduced by the switching protocol.
Hongwei and Ermira in the robot room.
back to topAutomation
Recently, we worked with two groups to develop robotic systems. The Universal Plate Pourer is an efficient, versatile instrument for filling a variety of media plates including round, rectangular and square formats. The BioMatrix is a system for manipulating high-density yeast arrays for a power user; it is high throughput, fully automated and can include stackers and cherry picking options.
back to top