Author: Zhao, Huabin; Ru, Binghua; Teeling, Emma C.; Faulkes, Christopher G.; Zhang, Shuyi; Rossiter, Stephen J.
Title: Rhodopsin Molecular Evolution in Mammals Inhabiting Low Light Environments Document date: 2009_12_16
ID: 02uqygfs_42
Snippet: Polymerase Chain Reactions (PCR) contained 1 ml (50 ng/ml) genomic DNA, 5 ml 10 x buffer, 1.5 ml (50 mM) MgCl 2 , 1 ml (10 mM) of each primer and 1 U Taq DNA polymerase (Takara). Reactions were performed on a DNA Engine Dyad Cycler (BioRad) with the following conditions: initial denaturation step of 5 min; 30 cycles of denaturation at 94uC for 30 s, annealing temperature (see Table S2 , Supplementary Material online) for 30 s; extension at 72uC f.....
Document: Polymerase Chain Reactions (PCR) contained 1 ml (50 ng/ml) genomic DNA, 5 ml 10 x buffer, 1.5 ml (50 mM) MgCl 2 , 1 ml (10 mM) of each primer and 1 U Taq DNA polymerase (Takara). Reactions were performed on a DNA Engine Dyad Cycler (BioRad) with the following conditions: initial denaturation step of 5 min; 30 cycles of denaturation at 94uC for 30 s, annealing temperature (see Table S2 , Supplementary Material online) for 30 s; extension at 72uC for 30 to 180 s (depending upon the target length), and a final extension of 72uC for 5 min. PCR products were checked on an agarose gel and cloned into a pMD19-T vector (Takara). Positive clones were sequenced on an ABI sequencer using the sequencing primer pair M13-47 and M13-48 (see Table S2 , Supplementary Material online). In order to avoid artifacts, multiple clones of each PCR product were sequenced in both forward and reverse directions.
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