BME100 s2017:Logistics

BME 100 Spring 2017	Home People Lab Write-Up 1 | Lab Write-Up 2 | Lab Write-Up 3 Lab Write-Up 4 | Lab Write-Up 5 | Lab Write-Up 6 Course Logistics For Instructors Photos Wiki Editing Help
COURSE SPECS ~100 Freshmen LAB WRITE-UP TEMPLATE PAGES Write-up 1 - Device Design Lab Write-up 2 - Device Design Lab Write-up 3 - Device Design Lab Write-up 4 - DNA Lab Write-up 5 - DNA Lab Write-up 6 - DNA Lab MATERIALS & METHODS - DNA AMPLIFICATION Reactions (100 μL total volume) contained: Template DNA - Kozak (BBa_BBa_J176012) inserted in plasmid vector V0120 (BBa_J176127) via standard BioBrick ligation; ~10 ng per "positive" reaction; negative reactions contained no plasmid Primers - Forward primer P0001 (5'-gggttttcccagtcacgacg), Reverse primer P0002 (5'-tgtggaattgtgagcggataaca). Amplicon size = 277 bp, spanning some of the vector, the BioBrick prefix sites, Kozak, the BioBrick suffix, and more vector sequence. 2x GoTaq colorless PCR Master Mix - Promega M7133 A few of the samples were checked via standard ethidium bromide-stained agarose gel electrophoresis outside of class (see [1]). All other samples were measured via SYBRgreen staining on an LED fluorimeter. All reactions were run on an Open PCR machine. MATERIALS & METHODS - DNA IMAGING SYBR Safe dye (Invitrogen) - 10,000x stock diluted to 1x in 1x TBE 100 μL PCR reaction was diluted in 500 μL buffer (1x TBE) to yield a 1/6 dilution of DNA 80 μL diluted DNA was applied to 80 μL SYBR Green dye directly on the hydrophobic slide on the fluorimeter (DNA is diluted 1/12 at this point) OBTAINING MATERIALS We have written several worksheets and hand-outs for this course. Please contact one of the instructors if you are interested in obtaining copies.