User:Karmella Haynes/Notebook/ASU courses/2014/01/24

ASU Courses

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01/24/14 Kozak plasmid minipreps Kozak Plasmid Minipreps For BME 100, spring 2014 Also for SynBERC Scholars/ Quanta "qOpenPCR" project Day 1 - Tasks for Behzad: Quick-transform Kozak miniprep into DH5α Grow on 100μg/mL amp plates overnight. Include zero plasmid negative control Day 2 - Tasks for Behzad Pick 4 colonies, innoculate 4 cultures, 5 ml each Grow overnight Day 3 - Tasks for Behzad Pellet the cells in the ancient table-top swing-bucket centrifuge Very thoroughly (by vortexing and flicking and pipetting up and down) resuspend cells in appropriate amount of water Proceed with miniprep Measure DNA concentrations

Colony	1	2	3	4
Conc.	30	227	243	45
260/280	1.9	1.87	1.9	1.9

Digest plasmids with restriction enzymes and gel electrophoresis (1% agarose) to confirm the assembled construct size

Plasmid DNA	3.0 μL
EcoRI	1.0 μL
PstI	1.0 μL
10x FastDigest buffer + green loading dye	1.5 μL
dH2O	8.5 μL
	15.0 μL total
Incubate at 37°C for 10 minutes.