The principle of 3' RACE (Rapid Amplification of cDNA Ends) would be used to isolate and clone the missing 3' end of the provided cDNA sequence. The following steps would be taken: 1) Design primers near the known 5' end of the cDNA sequence to amplify the unknown 3' region. 2) Use the primers to perform PCR on a cDNA library in order to amplify the full length cDNA. 3) Clone and sequence the amplified product to determine the complete cDNA sequence.

1. 2) Explain the principle behind 3 RACE. Describe in detail how you would use this technique to
isolate and clone the missing 3 end of this cDNA. (10 marks) Material at your disposal: everything
that youve accumulated so far in your genetic engineering toolbox. The sequence and optimization
of each primer must be detailed. taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaaag cggccgcgac tctagatcat aatcagccat
accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacct