Combined <i>RASSF1A</i> and <i>RASSF2A</i> Promoter Methylation Analysis as Diagnostic Biomarker for Bladder Cancer

<table class="table-group" id="tab2"><tr><td><table class="table"><tr><td class="thead-hr" colspan="7"><hr/></td></tr><tr class="thead"><td align="left"> Analysis</td><td align="center">Sense primer</td><td align="center">Antisense primer</td><td align="center">Product size (bp)</td><td align="center"><svg height="15.8375" id="M4" style="vertical-align:-3.39064pt" version="1.1" viewbox="0 0 43.575001 15.8375" width="43.575001" xmlns="http://www.w3.org/2000/svg" xmlns:xlink="http://www.w3.org/1999/xlink">
<g transform="matrix(.017,-0,0,-.017,.062,11.55)"><path d="M649 676l-22 -187l-33 -2q3 56 -12 94q-8 20 -31.5 26.5t-86.5 6.5h-74l-90 -491q-4 -23 -6 -36.5t1 -25t7 -16.5t18 -9t27 -5t41 -3l-6 -28h-286l4 28q68 5 84 18.5t28 76.5l94 491h-55q-74 0 -100.5 -6t-41.5 -23q-24 -29 -54 -98l-32 1q32 98 53 188h22q7 -18 15 -22
t37 -4h417q23 0 33.5 5t25.5 21h23z" id="x1D447"></path></g>
<g transform="matrix(.012,-0,0,-.012,11.388,15.637)"><path d="M433 39l-85 -51q-27 0 -47 20q-16 16 -24 44q-5 -3 -24 -16.5t-27 -18.5t-23 -13.5t-25 -12t-19 -3.5q-52 0 -86 36.5t-34 85.5q0 71 81 99q128 43 155 65v17q0 54 -23 83.5t-62 29.5q-28 0 -45 -19t-29 -66q-7 -23 -29 -23q-15 0 -29 13t-14 30t31 40q25 19 68.5 40.5
t81.5 29.5q49 0 82 -27q45 -39 45 -123v-185q0 -60 35 -60q18 0 36 11zM275 84v156q-12 -6 -49 -23t-41 -19q-31 -14 -46 -31t-15 -43q0 -35 22.5 -55.5t48.5 -20.5q19 0 42 10.5t38 25.5z" id="x61"></path></g><g transform="matrix(.012,-0,0,-.012,16.504,15.637)"><path d="M524 0h-209v26q44 5 55 18t11 65v167q0 114 -91 114q-50 0 -109 -51v-235q0 -50 10 -61.5t55 -16.5v-26h-217v26q51 5 62 17t11 61v206q0 47 -9.5 58.5t-50.5 19.5v23q82 15 139 40v-79l67 47q47 30 83 30q60 0 94.5 -40t34.5 -112v-193q0 -50 10 -61.5t54 -16.5v-26z
" id="x6E"></path></g><g transform="matrix(.012,-0,0,-.012,23.013,15.637)"><use xlink:href="#x6E"></use></g><g transform="matrix(.012,-0,0,-.012,29.474,15.637)"><path d="M380 106l15 -22q-42 -52 -87 -74t-76 -22q-89 0 -142 62t-53 151q0 103 66 175q67 73 146 73h1q61 0 105.5 -46.5t44.5 -105.5q0 -16 -4.5 -22.5t-18.5 -9.5q-54 -10 -262 -17q1 -95 46 -143.5t104 -48.5q61 0 115 50zM225 412q-35 0 -64.5 -32.5t-40.5 -90.5q83 0 170 3
q14 0 19 4.5t5 15.5q1 41 -23.5 70.5t-65.5 29.5z" id="x65"></path></g><g transform="matrix(.012,-0,0,-.012,34.59,15.637)"><use xlink:href="#x61"></use></g><g transform="matrix(.012,-0,0,-.012,39.873,15.637)"><path d="M238 0h-223v26q48 5 60 17t12 61v473q0 50 -10.5 60.5t-58.5 15.5v24q73 8 148 35v-608q0 -49 11.5 -61t60.5 -17v-26z" id="x6C"></path></g>
</svg> (°C)</td><td align="center">Unmethylation <svg height="15.8375" id="M5" style="vertical-align:-3.39064pt" version="1.1" viewbox="0 0 33.275002 15.8375" width="33.275002" xmlns="http://www.w3.org/2000/svg" xmlns:xlink="http://www.w3.org/1999/xlink">
<g transform="matrix(.017,-0,0,-.017,.062,11.55)"><use xlink:href="#x1D447"></use></g>
<g transform="matrix(.012,-0,0,-.012,11.388,15.637)"><path d="M797 0h-214v26q46 5 58 17.5t12 63.5v167q0 116 -88 116q-48 0 -112 -52q3 -18 3 -42v-195q0 -46 11 -58t53 -17v-26h-214v26q48 5 59.5 17t11.5 60v172q0 115 -88 115q-47 0 -109 -52v-235q0 -47 10 -59.5t48 -17.5v-26h-211v26q50 5 62.5 17t12.5 60v207q0 48 -9 59.5
t-52 18.5v23q79 14 139 41v-81q3 2 29 20.5t44 29.5q45 28 81 28t66 -22t46 -61q25 20 83 57q39 26 76 26q59 0 93 -40t34 -113v-193q0 -49 10.5 -60.5t54.5 -16.5v-26z" id="x6D"></path></g><g transform="matrix(.012,-0,0,-.012,21.085,15.637)"><use xlink:href="#x65"></use></g><g transform="matrix(.012,-0,0,-.012,26.094,15.637)"><use xlink:href="#x6C"></use></g><g transform="matrix(.012,-0,0,-.012,28.962,15.637)"><path d="M298 36l-68 -35q-25 -13 -42 -13q-98 0 -98 119v290h-69l-4 11l28 29h45v72l65 67l14 -2v-137h113q8 -8 6 -21t-12 -19h-107v-256q0 -96 59 -96q29 0 61 17z" id="x74"></path></g>
</svg> (°C)</td><td align="center">Methylation <svg height="15.8375" id="M6" style="vertical-align:-3.39064pt" version="1.1" viewbox="0 0 33.275002 15.8375" width="33.275002" xmlns="http://www.w3.org/2000/svg" xmlns:xlink="http://www.w3.org/1999/xlink">
<g transform="matrix(.017,-0,0,-.017,.062,11.55)"><use xlink:href="#x1D447"></use></g>
<g transform="matrix(.012,-0,0,-.012,11.388,15.637)"><use xlink:href="#x6D"></use></g><g transform="matrix(.012,-0,0,-.012,21.085,15.637)"><use xlink:href="#x65"></use></g><g transform="matrix(.012,-0,0,-.012,26.094,15.637)"><use xlink:href="#x6C"></use></g><g transform="matrix(.012,-0,0,-.012,28.962,15.637)"><use xlink:href="#x74"></use></g>
</svg> (°C)</td></tr><tr><td class="thead-hr" colspan="7"><hr/></td></tr><tr><td align="left"><i>RASSF1A</i></td><td align="center">ATGTTAAGGGAATTTATTTAGAATGTATTT</td><td align="center">AACCTTCACTTAAAATAAAAAAAA</td><td align="center">146</td><td align="center">53</td><td align="center">73.2</td><td align="center">75.6</td></tr><tr><td align="left"><i>RASSF2A</i></td><td align="center">GAAGGGGTAGTTAAGGGGTAG</td><td align="center">CCTCTACTCATCTATAACCCAAATAC</td><td align="center">176</td><td align="center">55</td><td align="center">76.6</td><td align="center">81</td></tr><tr><td align="left"><i>RASSF4</i></td><td align="center">TAAATGGTTTGTGGTTTTTTGTTTT</td><td align="center">AAAAACACCTTTATACAATCTAACC</td><td align="center">175</td><td align="center">55</td><td align="center">72.8</td><td align="center">74</td></tr><tr><td align="left"><i>RASSF5</i></td><td align="center">GAAAGAGGTAGGGTTGAAGGTTTAG</td><td align="center">TCACCTAAAACAACTACAAAATTCC</td><td align="center">105</td><td align="center">55</td><td align="center">74</td><td align="center">75.6</td></tr><tr><td align="left"><i>RASSF6</i></td><td align="center">GTTTAGTTGAGTTATGTTTTGGGAG</td><td align="center">AAAAAACCAATACCCTATCTCTACC</td><td align="center">126</td><td align="center">55</td><td align="center">75.8</td><td align="center">79.2</td></tr><tr class="table-tr"><td colspan="7"><hr class="tbody-hr"/></td></tr></table></td></tr><tr class="table-fn"><td><svg height="15.8375" id="M7" style="vertical-align:-3.39064pt" version="1.1" viewbox="0 0 43.575001 15.8375" width="43.575001" xmlns="http://www.w3.org/2000/svg" xmlns:xlink="http://www.w3.org/1999/xlink">
<g transform="matrix(.017,-0,0,-.017,.062,11.55)"><use xlink:href="#x1D447"></use></g>
<g transform="matrix(.012,-0,0,-.012,11.388,15.637)"><use xlink:href="#x61"></use></g><g transform="matrix(.012,-0,0,-.012,16.504,15.637)"><use xlink:href="#x6E"></use></g><g transform="matrix(.012,-0,0,-.012,23.013,15.637)"><use xlink:href="#x6E"></use></g><g transform="matrix(.012,-0,0,-.012,29.474,15.637)"><use xlink:href="#x65"></use></g><g transform="matrix(.012,-0,0,-.012,34.59,15.637)"><use xlink:href="#x61"></use></g><g transform="matrix(.012,-0,0,-.012,39.873,15.637)"><use xlink:href="#x6C"></use></g>
</svg>, primer annealing temperature<br/><svg height="15.8375" id="M8" style="vertical-align:-3.39064pt" version="1.1" viewbox="0 0 33.275002 15.8375" width="33.275002" xmlns="http://www.w3.org/2000/svg" xmlns:xlink="http://www.w3.org/1999/xlink">
<g transform="matrix(.017,-0,0,-.017,.062,11.55)"><use xlink:href="#x1D447"></use></g>
<g transform="matrix(.012,-0,0,-.012,11.388,15.637)"><use xlink:href="#x6D"></use></g><g transform="matrix(.012,-0,0,-.012,21.085,15.637)"><use xlink:href="#x65"></use></g><g transform="matrix(.012,-0,0,-.012,26.094,15.637)"><use xlink:href="#x6C"></use></g><g transform="matrix(.012,-0,0,-.012,28.962,15.637)"><use xlink:href="#x74"></use></g>
</svg>,  amplicon melting temperature.<br/></td></tr></table>

Primer sequences and melting temperature for <i>RASSF</i> genes.

Molecular Biology International

tab2

Table 2

Table 2: Combined <i>RASSF1A</i> and <i>RASSF2A</i> Promoter Methylation Analysis as Diagnostic Biomarker for Bladder Cancer 

Molecular Biology International

Combined RASSF1A and RASSF2A Promoter Methylation Analysis as Diagnostic Biomarker for Bladder Cancer

Table 2