Team:Cornell/testing/project/wetlab/4/3

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<h3>Napthalene</h3>
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<h3>RT-qPCR</h3>
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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:
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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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Revision as of 01:39, 4 October 2012

qPCR

RT-qPCR

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:

FOR_enzA: CAGCCTTTTACCCAAGGTGA
REV_enzA: CACGATTCGAGAGGGTGATT
FOR_RecA: TTCCCCTCGACATTGTCATCATCGGA
REV_RecA: AAGGGCGATAAAATTGGTCAAGGCCG
3'_FOR_qPCR_mtrB: ACGCTCAATATCAAGCCACCGAGA
3'_REV_qPCR_mtrB: TGTGCGGTGTAGTCATGGCTGT

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