Design two degenerate primer sequences

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Reference no: EM1383719

Here is the amino acid sequence of the enzyme (in single letter code):

 

MAAMDMDQFLAAAIDAAKKAGQIIRKGFYETKHVEHKGQVDLVTETDKGCEELVFNHLKQLFPNHKFIGEETTAAFGVTELTDEPTWIVDPLDGTTNFVHGFPFVCVSIGLTIGKVPVVGVVYNPIMEELFTGVQGKGAFLNGKRIKVSAQSELLTALLVTEAGTKRDKATLDDTTNRINSLLTKVRSLRMSGSCALDLCGVACGRVDIFYELGFGGPWDIAAGIVIVKEAGGLIFDPSGKDLDITSQRIAASNASLKELFAEAMMLTAA

 

Design two degenerate primer sequences that will amplify the entire Open reading frame (ORF) of the corresponding gene in a PCR reaction.  To facilitate the cloning of the gene after PCR amplification, place a restriction endonuclease site at each end of your primers.  You may choose any restriction endonuclease site you like, or use EcoRI (GAATTC).  Hints: restriction endonuclease sites are cleaved more efficiently when 1-2 extra nucleotides are included at each end; these extra nucleotides are usually Gs or Cs. You need a forward and a reverse primer.  In general, primers that are 18-30 nucleotides long are suitable for PCR amplification.  Write out your primer sequences 5’ to 3’.

 

 

1. Forward primer sequence:

 

 

 

 

 

 

 

2. Reverse primer sequence:

Reference no: EM1383719

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