a. Design a primer pair, each 18-40 nucleotides long, to amplify the underlined sequence.
Keep in mind this is only one strand in the 5′-3′ orientation, you need to provide the 3′-5′ complementary prior to designing your primers. Both primers cannot be made on one strand, forward is on one strand, reverse is on the other.
5′-CCTCCGATCT CCAGCTGGAG CGTATGAACG TCTACTTCAA CGAGGTGCGTACCTCAAAAT TTCAGCATCT ATGAAAACGC TTTGCAACTC CTGACCGCTTCTCCAGGCCA GCGGAAACAA GTATGTCCCT CGTGCCGTCC TACTCTCCTGATCTCCAAGA TCCGTGAGGA GTTCCCCGAC CGTATGATGG CCACCTACTCCGTTGTCCCC TCCC CCAAGG TCTCCGACAC CGTTGTTGAG-3′
Primer 5′ – 3′ Sequence
Forward
Reverse
b. Optimize your primers.
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