The amplification efficiency of foraminifera DNA differs based on different PCRprimers. The improper primers can even cause the failure of PCR amplificationinmolecular experiment,which will lead tomisleading result.In order tofind themostsuitablePCR primers ofbenthicforaminiferaof18S rDNA(18SRinbosomalRNA)in different sediment types of diverse marine habitats,the research collected the surfacesediments of 15 station in Qingdao Bay, Jiaozhou Bay, Yellow Sea, Northwest Pacificand East Pacific Ocean.The DNA of foraminiferawas extracted and we useddifferentforaminifera-specific primersto amplify DNA ofbenthic foraminiferain these areas.By comparingthe Gel electrophoresisandthegrayscale value calculated by Image Jsoftware, the result showed thatbenthic foraminifera in various sediments havethemost suitableforaminifera-specific primers.In order toexplore the regular patternthatusing different foraminifera-specificin different sediment sea areas,we conductedmoleculardiversity analysis of the benthic foraminifera to obtain the communitycomposition ofthesein different sediments.The workin this studyincluded:
Explore effective PCR primersofbenthic foraminiferain various sediment
By comparing theGel electrophoresis and the grayscale value calculated by Image Jsoftware,the results showed that s14F3 (forward primer)/s17 (reverse primer) wereeffective for intertidal sediments; s14F1 (forward primer)/s17 were more effective forshallow coastal sediments; s14F3/s15ROTEX (reverse primer) had a betteramplification effect on the offshore shelf sediments; s14F3/s17 exhibited better effectsfor deep-sea muddy sediments; s14F1/s15ROTEX were effective for metal noduledeposits.
The analysis of molecular diversityabout benthic foraminifera in different areas
The result suggested that there are many vitreous foraminiferain the Qingdao Bay,Jiaozhou Bay and theEast Pacific. However, the benthic foraminifera of this areamainlyconsist ofagglutinated foraminifera. In addition, it indicated that the water depth of sediment and the sediment itself(biological and non-biological) as well as the composition of foraminifera in thesedimentsplayedan importantrole inthechoice of primers of benthic foraminifera.Thus, the appropriate primers would improve the amplification efficiency of PCR forforaminifer from different sediments. Our study will provide an effective technicalmethod for studies involving the molecular diversity ofbenthic foraminifera in differentmarine sediment types.
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