The mitochrondrial mutS (msh1) locus was used for DNA barcoding because it is a commonly used locus unique to corals65 , allowing for clean amplification even in the presence of zooxanthellae. A single coral polyp was macerated in 50 μL dimethyl sulfoxide, which was used at a concentration of 1% in a PCR using primers 5’-GCCATTATGGTTAACTATTAC-’366 and 5’-TSGAGCAAAAGCCACTCC-3’67 , and 5Prime HotMasterMix polymerase (Quanta Bio) according to the manufacturer’s instructions. The amplicon was Sanger sequenced using the former primer and verified to correspond to C. imbricata using NCBI MOLE-BLAST. For the sequence see Supplementary Note 2 .
5prime hotmastermix polymerase
Manufactured by Quantabio
The 5Prime HotMasterMix polymerase is a high-performance PCR reagent that enables rapid and efficient DNA amplification. The polymerase enzyme exhibits robust thermal stability and exhibits enhanced processivity, allowing for reliable and consistent results.
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2 protocols using 5prime hotmastermix polymerase
1
Coral DNA Barcoding Using msh1 Locus
2
Coral DNA Barcoding Using msh1 Locus
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The mitochrondrial mutS (msh1) locus was used for DNA barcoding because it is a commonly used locus unique to corals65 , allowing for clean amplification even in the presence of zooxanthellae. A single coral polyp was macerated in 50 μL dimethyl sulfoxide, which was used at a concentration of 1% in a PCR using primers 5’-GCCATTATGGTTAACTATTAC-’366 and 5’-TSGAGCAAAAGCCACTCC-3’67 , and 5Prime HotMasterMix polymerase (Quanta Bio) according to the manufacturer’s instructions. The amplicon was Sanger sequenced using the former primer and verified to correspond to C. imbricata using NCBI MOLE-BLAST. For the sequence see Supplementary Note 2 .
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