Supplemental File S6: Answer key for the example worksheets to assess achievement of learning
objectives 3, 4, and 6.
Instructions
1. Using the DNA template shown below, underline the template sequence where the two primers will annel.
Label the underlined sequence as “Primer 1” or “Primer 2”.
2. Using the DNA template and primers shown below, write the sequence of the resulting PCR product (after
30 cycles). Be sure to align your product with the sequence shown.
3. Draw a box around the site(s) on the PCR product that would be recognized and cleaved by EcoRI.
(3’5’ Primer 2)
3-‘gcatattgccccatc-5’
5’-GTTGTATCGTACATAGGATCGATAGGCTAGACGAATTAGACTTACGTATAACGGGGTAGACAGATATTTAG-3’
3’-CAACATAGCATGTATCCTAGCTATCCGATCTGCTTAATCTGAATGCATATTGCCCCATCTGTCTATAAATC-5’
5’-tcgtacataggatcg-3’
Primer 1 (5’3’)
Primer 1:
Primer 2:
5’ CGAATTCTCGTACATAGGATCC 3’
5’ GGAATTCCTACCCCGTTATACG 3’
EcoRI recognition site:
5’-GAATTC-3’
3’-CTTAAG-5’
5’-CGAATTCTCGTACATAGGATCGATAGGCTAGACGAATTAGACTTACGTATAACGGGGTAGGAATTCC-3’
3’-GCTTAAGAGCATGTATCCTAGCTATCCGATCTGCTTAATCTGAATGCATATTGCCCCATCCTTAAGG-5’
Instructions
The diagram below shows a 310 bp fragment of double-stranded DNA. Above the fragment, the sizes of
various segments of DNA are shown (in base pairs). Below the DNA fragment, various primers, labeled A
through F are shown.
30
Size
(in bp)
50
30
150
30
20
5’
3’
5’
3’
1.
3’
5’
A
B
3’
5’
5’
3’
C
D
3’
5’
5’
3’
E
F
3’
5’
Predict the size of the PCR product (if any) that will be produced from the following primer pairs.
Primers
Product? yes/no
A and CNn
no
A and F
yes
C and E
no
D and F
no
Product Size (bp)
290
2.
Which primer pairs will produce a product that is 210 bp? Primers C and F
3.
Which primer pairs will produce a product that is 190 bp? Primers A and D