Team:Cornell/project/wetlab/results/qPCR

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					<h6>Wet Lab</h6>
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					<a href="https://2012.igem.org/Team:Cornell/project/wetlab">Overview</a>
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					<a href="https://2012.igem.org/Team:Cornell/project/wetlab/chassis">Chassis</a>
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					DNA Assembly
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							<a href="https://2012.igem.org/Team:Cornell/project/wetlab/assembly/arsenic">Arsenic Reporter</a>
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							<a href="https://2012.igem.org/Team:Cornell/project/wetlab/assembly/salicylate">Salicylate Reporter</a>
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					Testing &amp; Results
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							<a href="https://2012.igem.org/Team:Cornell/project/wetlab/results/fluorescence">Fluorescence</a>
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							<a href="https://2012.igem.org/Team:Cornell/project/wetlab/results/qPCR">qPCR</a>
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							<a href="https://2012.igem.org/Team:Cornell/project/wetlab/results/western">Western</a>
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							<a href="https://2012.igem.org/Team:Cornell/project/wetlab/results/artificial_river_media">Artificial River Media Growth Assays</a>
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							<a href="https://2012.igem.org/Team:Cornell/project/wetlab/results/naphthalene_growth_assays">Naphthalene Growth Assays</a>
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					<a href="https://2012.igem.org/Team:Cornell/project/wetlab/future_work">Future Work</a>
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					<h2 class="centered">qPCR</h2>
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					<h3>RT-qPCR</h3>
We are currently using two-step RT-qPCR to confirm that our engineered Shewanella strains respond to arsenic and naphthalene. Total RNA will be isolated using the E.Z.N.A.™ Bacterial RNA Isolation Kit from Omega bio-tek; cDNA will be synthesized using the qScript™ Flex cDNA Kit from Quanta Biosciences; and we are using the ABI ViiA7 platform along with the KAPA SYBR(R) FAST qPCR Kit from KAPA Biosystems. Our primers are as follows: 
<br><br>
FOR_enzA: CAGCCTTTTACCCAAGGTGA
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REV_enzA: CACGATTCGAGAGGGTGATT
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FOR_RecA: TTCCCCTCGACATTGTCATCATCGGA
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REV_RecA: AAGGGCGATAAAATTGGTCAAGGCCG
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3'_FOR_qPCR_mtrB: ACGCTCAATATCAAGCCACCGAGA
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3'_REV_qPCR_mtrB: TGTGCGGTGTAGTCATGGCTGT
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