1a). Add 20 nucleotides of coding sequence to the following forward and reverse

Question

1a). Add 20 nucleotides of coding sequence to the following forward and reverse primers. the nucleotides necessary for optimal digestion of the amplicon are italized while the restriction enzyme recognition sites are represented in bold type)

b). What will be the theoretical length of the expected PCR product to be amplified using your section primers? Specify the number of nucleotides. Be as specific as possible in calculating the expected length: Indicate the exact number of nucleotides that will be included in the amplicon.

SLOHCDDFRE HRPAYRGPDSYSSVIASHEPCDPEEMTRVWNECRSANYA ORIGIN 1 ctgtaaaaca attttttaga tttgtaagca ctcaatttga atgcttttag tcttagttat 61 tttatttttc gtacggtaga tgggcgttta gtattttaac cgcatcacgg tcacactggc 9 121 ggatttgttt ttattttggt cgcc tcat cettct ctqaact taga cttcactac gaacg atgacactgg a a gttgace agectgegeg getcacgatg ctcataaaga gegeggtggg caattegeg ccattc 45..1122 /gene-"br /locus_tag- "Dmel CG4934" /gene synonyn "brc; Brn; CG4934; DmelICG4934 EG:EG0007.6 fs (1)107; fs (1)A107; 1(1)6P6 /EC number=" 2-4·1·- · note- "CG4934 gene product fron transcript cG4934-RA; CG4934-PA; brn-PA; gene B /codon start l /product-"brainiac" /protein id- NP 476901.1 /db_xref "FLYBASE: FBpp0070606 /db_xref- "GencID: 3135 /db xref- "FLYBASE: Fagn0000221" /translation- "MOSKHRKLLLRCLLVLPLILLVDYCGLLTHLHELNFERHFHYPL NDDTGSGSASSGLDKFAYLRVPSETAEVPVDQPARLTMLIKSAVGNSRRREAIRRTWG agacattet qca ct atcagttca atc a gttctatctt ttegtggacg acgactacta tgttteggce aagaacgtge teaagtttet gggcaggggg at acccettcga tc cttccggttc aagcgggca ttteccttea gcactgcgac gactteeget tecateggce ggegtacaaa acqaat to atec cqaçcagat acgcgggtat ggaacga aattac agcaaga acaggcagga 1141 ctgaagageg ctgeagattg tacatatata cgaacgagac agcacctaga caacagcctt 1201 ataaagtata ttaggagcag tggatgatac ctcaaccgtt cagtgagact ttaaaccaag 1261 cagetggtge taatagtgea agatcgcaag agtaattcaa ttctaattgt aagegtaatg 1321 ataaagcagt tgattcaatt cgtatatata gagattatgt gcaaatgtcg gataacgtat 1381 gtgtggtctg atacctttga atgcaatata aagttgatta acataggaaa gcgaataaad 1441 aacttaaaat ASDOFNRSEFYLFVDDDYYVSAKNVLKFLGRGROSHOPELLFAGHVFOTSPLRHRFSR NYVSLEEYPFDRMPPYVTAGAFILSOKALROLYAASVHLPLFRFDDVYLGIVALKAGI SLOHCDDFRFHRPAYKGPDSYSSVIASHEEGDPEENTRVNNECRSANYA

Explanation / Answer

I believe the correct answers to be:

A) Forward Primer : 5'-GTGTATAAGCTTATGCAAAGTAAACACCGCAA

Reverse Primer: 5'-ATTTATGGTACCCTACGCGTAATTGGCGGATC

B) The amplicon as shown in the figure and primer included should be of 1000 bp in total.

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