- ~230 Freshmen
- 3 Lab sections, ~85 students per section
- 10 - 17 groups, of ~5 students per group
- 3 hours per lab
MATERIALS & METHODS - DNA AMPLIFICATION
- Reactions (100 μL total volume) contained:
- Template DNA - Kozak (BBa_BBa_J176012) inserted in plasmid vector V0120 ([http://partsregistry.org/wiki/index.php?title=Part:BBa_J176127 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 ). 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
- We have written several worksheets and hand outs for this course.
- Please contact one of the instructors if you are interested in obtaining copies.