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Production of a cDNA library to Harmonia Axyridis Multicolor Lady Beetle

      RATIONALE: Sensitivity to Harmonia Axyridis also known as multicolor lady beetle has been described previously and two allergens having a molecular weight of 16.6 and 30 KD has been identified.
      METHODS: In this study we sought to isolate mRNA from the beetles to prepare a cDNA library. One gram of beetles was frozen in liquid nitrogen and pulverized. The resulting material was extracted in Guanidine Thiocyanate and separated by ultra centrifugation on a Cesium Chloride gradient. The resulting total rRNA pellet was resuspended and extracted in phenol:chloroform.
      RESULTS: The RNA was quantitated and the mRNA extracted over an Oligo(dt) column. Using Promega (Madison, WI) Universal RiboClone cDNA Synthesis System a cDNA library was prepared from the extracted mRNA.
      CONCLUSIONS: We concluded the cDNA library has hundreds of clones containing the complete cDNA sequences of the identified allergens. Further work is necessary to determine which of these are producing the allergens.