BME100 s2017:Logistics

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BME 100 Spring 2017 Home
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Lab Write-Up 1 | Lab Write-Up 2 | Lab Write-Up 3
Lab Write-Up 4 | Lab Write-Up 5 | Lab Write-Up 6
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COURSE SPECS

  • ~100 Freshmen



LAB WRITE-UP TEMPLATE PAGES



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.



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.