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=== <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">mRNA as vaccine for animal study</span> === <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">Template preparation by PCR using 2X Taq PCR mix</span> {| border="1" cellspacing="1" cellpadding="1" style="width: 577px;" |- | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">Clone #</span><br/> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">Name</span> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">Des</span> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">F Primer</span><br/> | style="width: 165px;" | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">R Prime</span> | style="width: 37px;" | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">PCR Length</span> |- | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">2684</span><br/> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">DS2</span> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">T7-IRES-DS2</span> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">T7-IRES-F</span> <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">or T7-F</span> <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">or IVT-T7-F</span> | style="width: 165px;" | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">Flag-R</span><br/> | style="width: 37px;" | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">997</span> |- | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">2838</span> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">DS2</span> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">T7-DS2 in pJet</span> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">T7-DS2-F</span><br/> | style="width: 165px;" | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">Flag-R</span> | style="width: 37px;" | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">454</span> |- | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">2831</span><br/> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">DS2-fd</span> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">T7-DS2-fd in pJet</span> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">T7-DS2-F</span><br/> | style="width: 165px;" | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">Foldon-R</span><br/> | style="width: 37px;" | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">500</span> |- | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">2823</span> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">DS2</span> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">T7-DS2 in pJet</span><br/> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">T7-DS2-F</span><br/> | style="width: 165px;" | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">Flag-R</span><br/> | style="width: 37px;" | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">454</span><br/> |- | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">2823</span> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">DS2 <span style="color: rgb(0, 0, 0); font-family: Arial, Verdana, sans-serif; font-size: 12px;">SAM</span></span> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">T7-DS2 in pT7-VEE</span><br/> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">T7 or T7-VEE-F</span> | style="width: 165px;" | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">Flag-R or VEE-pA-R</span><br/> | style="width: 37px;" | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">8025</span> |- | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">2831</span> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">DS2-fd SAM</span> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">T7-DS2-fd in pT7-VEE</span> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">T7 or T7-VEE-F</span> | style="width: 165px;" | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">Foldon-R or VEE-pA-R</span><br/> | style="width: 37px;" | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">8065</span> |- | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">2874</span><br/> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">RBD-fd</span> | <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">Full length RBD fuse to fd, with polyA</span> | <br/> | style="width: 165px;" | <br/> | style="width: 37px;" | <br/> |} <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">Flag-R TTTTTTTTTTTTTTTCTCGAGTTACTTGTCGTCATCGTCTTTG</span> <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">(CAAAGACGATGACGACAAGTAACTCGAGAAAAAAAAAAAAAAA)</span> <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">T7-DS2-F TAATACGACTCACTATAAGGAGGCCACCATGGTCAGACAAATC</span> <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">Foldon-R tttttttttttttttttttttacaagaaggtcgacagcag</span> <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">(ctgctgtcgaccttcttgtaaaaaaaaaaaaaaaaaaaaa)</span> <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">VEE-pA-R GAATTAATTCCCCTCGAcgcgttctag</span> <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">T7-VEE-F T<u>AATACGACTCACTATA</u>''<span style="color:#FF0000;">'''A'''</span>G''ATGGGCGGCGCATGAGAG</span> <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">After PCR, take 5 ul to check purity on agarose gel. Purify the remaining PCR product using a spin column, elute DNA using 25 ul ddH2O, measure DNA concentration using nanodrop</span> <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">[[File:DS2 DS2fd PCR.JPG|RTENOTITLE]]</span> <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">[[File:2838 2831 PCR1.JPG|RTENOTITLE]]</span> <span style="color: rgb(37, 37, 37); font-family: sans-serif; font-size: 14px;">[[File:2838 2831 PCR2.JPG|RTENOTITLE]]</span>
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