You want to amplify by PCR the DNA encoding the CDS that is the second from the 5’ end of the fragment (in the direction it was given to you), From START to the STOP codon . Design the primers that will you use to amplify this fragment post their sequences, specifying the 5’ and 3’ ends and the Tm. Post a picture showing where the oligonucleotide will hydridize on the sequence.
Once you have amplified the PCR product you want to check it by digesting it with AccI. How many DNA fragments will you see on the agarose gel and what are their sizes in base pair. Upload the picture of the virtual digest.
Protein Sequence of CDS: MKITPIKTRKVFAHDSLQEILQEALPPLQERSVVVVSSKIVSLCEGAVADARMCKAELIKKEADAYLFCE KSGIYLTKKEGILIPSAGIDESNTDQPFVLYPKDILGSCNRIGEWLRNYFRVKELGVIITDSHTTPMRRG VLGIGLCWYGFSPLHNYIGSLDCFGRPLQMTQSNLVDALAVAAVVCMGEGNEQTPLAVIEQAPNMVYHSH PTSREEYCSLRIDETEDLYGPFLQAVTWSQEKK
Below is the 3'5' Frame 3 with the CDS above PRSTQSCKFQHK-RGAMTSWNFVCLSLGSNLGNRHEHIRRAYASLKKAGIRNLKSSVILE TKALLLEGAPKEWDLPYFNSVVIGETQLSPDELIEEIKMIESRFGQDASLKWGPRPIDID VLFYGDEAFSYHSDKCTIPHPKVLERPFLLSMIASLCPYRRFRLEGSSCNGKTFAELAAI YPLTEEDALGSFGSATQIMGIVNITDNSISDTGLFLEARRAAAHAERLFAEGASIIDLGA QATNPRVKDLGSVEQEWERLEPVLRLLAERWGAAQQCPDVSIDTFRPEIIRRAVEVFPIR WINDVSGGSLEMAHLAKEFGLRLLINHSCSLPPRPDCVLSYEESPIEQMLRWGESQLEQF AQVGLDTSWQVVFDPGIGFGKTPVQSMLLMDGVKQFKRVLECPVLIGHSRKSCLSMLGRF NSNDRDWETIGCSVSLHDRGVDYLRVHQVEGNRRALAAAAWAGMFV-SKQQVSLLLIPEE -WEL-ASSLGVIPKIYVFLQKPFEIIPSLWDERLGSLFQTSISMGGISLSFLAGCIHHNA -EFLPLQSMGHYL-IIRF-LGERSSLKVFSNKTF-KLVLSHISKRNIGAILSSLSRDYQD GRRNVFVIQRISVFIIMKITPIKTRKVFAHDSLQEILQEALPPLQERSVVVVSSKIVSLC EGAVADARMCKAELIKKEADAYLFCEKSGIYLTKKEGILIPSAGIDESNTDQPFVLYPKD ILGSCNRIGEWLRNYFRVKELGVIITDSHTTPMRRGVLGIGLCWYGFSPLHNYIGSLDCF GRPLQMTQSNLVDALAVAAVVCMGEGNEQTPLAVIEQAPNMVYHSHPTSREEYCSLRIDE TEDLYGPFLQAVTWSQEKK-WRCL-IF-IS-I-LFKISIC-NTHFM-NGRRGSLLKSNYR RMPKTIIYISKPFLNIYLRFIVVAMI-RRVSYC-IT-WMKRTVTLIILICGSSLCLL-EL LQKS-RLMSLVKQQKRK-LLSCGGVQEIL-LQEWLLCILMRVKFHVSLERKFVD-LSTLD FPILKTMHISQNMKKRMCGMLEKKKRSLRCFSKMTLIKC-RHRKK-RNLCMAF-ILFWIQ ELVVVVINLIKMI-QLLSA
Hi tiffspla001,
This community of volunteers isn't here to solve your homework questions. If you are working on this and got stuck somewhere we would be happy to help, but show some effort from your side and tell us what you tried before just copy pasting the question here.
Cheers, Wouter
Actually, I was just looking for help about how I should go about doing this, since I really don't even know where to begin.
You probably had some lessons about this?
This looks like a homework than a doubt.
Actually, I was just looking for help about how I should go about doing this, since I really don't even know where to begin.