BackgroundTo avoid destructive sampling for conservation and genetic assessment, we isolated the DNA of clam Cyclina sinensis from their feces. DNA electrophoresis and PCR amplification were used to determine the quality of fecal DNA. And we analyzed the effects of different conditions on the degradation of feces and fecal DNA.ResultsThe clear fecal DNA bands were detected by electrophoresis, and PCR amplification using clam fecal DNA as template was effective and reliable, suggesting that clam feces can be used as an ideal material for noninvasive DNA isolation. In addition, by analyzing the effects of different environmental temperatures and soaking times on the degradation of feces and fecal DNA, we found that the optimum temperature was 4 °C. In 15 days, the feces maintained good texture, and the quality of fecal DNA was good. At 28 °C, the feces degraded in 5 days, and the quality of fecal DNA was poor.ConclusionsThe clam feces can be used as an ideal material for noninvasive DNA isolation. Moreover, the quality of fecal DNA is negatively correlated with environmental temperature and soaking time.
The reversible black shell colour change in the clam Cyclina sinensis functions as an adaptation to different environmental conditions (within or outside of muddy sediment) due to changes in the melanin redox state in the shell. The microphthalmia‐associated transcription factor (MITF) is a melanocyte‐specific transcription factor that is involved in the differentiation, proliferation and survival of melanocytes. In this study, the full‐length cDNA sequence of the mitf‐like gene was obtained from C. sinensis using RACE PCR. The mitf‐like cDNA is 1725 bp in length, containing a 47 bp 5′ untranslated region (UTR), a 1398 bp open reading frame and a 280 bp 5′ untranslated region. The mitf‐like gene contains seven exons separated by six introns, and it encodes a putative protein of 465 amino acids with a highly conserved domain of MITF containing a basic helix‐loop‐helix leucine zipper (bHLH‐LZ). The phylogenetic analysis of amino acid sequences using neighbour‐joining and maximum likelihood methods revealed that C. sinensis is closely related to Meretrix petechialis. Additionally, the mRNA expression of mitf‐like and melanin was high in the outer margin of the mantle, suggesting that the mitf‐like might play an important role in melanin synthesis in the mantle by regulating the cell cycle and survival of melanocytes; however, its role in C. sinensis shell melanin deposition requires further investigation.
Background
To avoid destructive sampling for conservation and genetic assessment, we isolated the DNA of clam Cyclina sinensis from their feces. DNA electrophoresis and PCR amplification were used to determine the quality of fecal DNA. And we analyzed the effects of different conditions on the degradation of feces and fecal DNA.Results
The clear fecal DNA bands were detected by electrophoresis, and PCR amplification using clam fecal DNA as template was effective and reliable, suggesting that clam feces can be used as an ideal material for noninvasive DNA isolation. In addition, by analyzing the effects of different environmental temperatures and soaking times on the degradation of feces and fecal DNA, we found that the optimum temperature was 4 °C. In 15 days, the feces maintained good texture, and the quality of fecal DNA was good. At 28 °C, the feces degraded in 5 days, and the quality of fecal DNA was poor.Conclusions
The clam feces can be used as an ideal material for noninvasive DNA isolation. Moreover, the quality of fecal DNA is negatively correlated with environmental temperature and soaking time.
Background
To avoid destructive sampling for conservation and genetic assessment, we isolated the DNA of clam Cyclina sinensis from their feces. DNA electrophoresis and PCR amplification were used to determine the quality of fecal DNA. And we analyzed the effects of different conditions on the degradation of feces and fecal DNA.Results
The clear fecal DNA bands were detected by electrophoresis, and PCR amplification using clam fecal DNA as template was effective and reliable, suggesting that clam feces can be used as an ideal material for noninvasive DNA isolation. In addition, by analyzing the effects of different environmental temperatures and soaking times on the degradation of feces and fecal DNA, we found that the optimum temperature was 4 °C. In 15 days, the feces maintained good texture, and the quality of fecal DNA was good. At 28 °C, the feces degraded in 5 days, and the quality of fecal DNA was poor.Conclusions
The clam feces can be used as an ideal material for noninvasive DNA isolation. Moreover, the quality of fecal DNA is negatively correlated with environmental temperature and soaking time.
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