Fluorescent Cell Sorting and Screening

All restriction endonucleases, Q5 polymerase, and Gibson assembly HiFi master mix were from New England Biolabs, Inc. GOTaq Flexi DNA polymerase was from Promega. Primers were obtained from Integrated DNA Technologies (IDT). A Sony MA900 fluorescence activated cell sorter (FACS) was used for sorting nonfluorescent cells from fluorescent cells, and an Azure C600 imager was used for screening of fluorescent colonies as previously described (5 (link)).

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Lal P.B., Wells F., Myers K.S., Banerjee R., Guss A.M, & Kiley P.J. (2021). Improving Mobilization of Foreign DNA into Zymomonas mobilis Strain ZM4 by Removal of Multiple Restriction Systems. Applied and Environmental Microbiology, 87(19), e00808-21.

Publication 2021

Q5 polymerase Gibson assembly HiFi master mix GOTaq Flexi DNA polymerase

Azure Cells Dna polymerase Fluorescence Primers Promega Restriction endonucleases

Corresponding Organization :

Other organizations : Great Lakes Bioenergy Research Center, University of Wisconsin–Madison, Oak Ridge National Laboratory

Top 5 similar protocols

Variable analysis

independent variables

Restriction endonucleases
Q5 polymerase
Gibson assembly HiFi master mix
GOTaq Flexi DNA polymerase
Primers

dependent variables

Fluorescent cells
Nonfluorescent cells

control variables

New England Biolabs, Inc. as the source for restriction endonucleases, Q5 polymerase, and Gibson assembly HiFi master mix
Promega as the source for GOTaq Flexi DNA polymerase
Integrated DNA Technologies (IDT) as the source for primers
Sony MA900 fluorescence activated cell sorter (FACS) for sorting nonfluorescent cells from fluorescent cells
Azure C600 imager for screening of fluorescent colonies

positive controls

None specified

negative controls

None specified

Annotations

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