User:David Johnston Monje/Notebook/Maize Endophyte Biofertilizers/2013/06/20

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Applied Soil Microbial Ecology Main project page
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Remains of the Day

  • Gonna combine optimized Oomycete DC6 primer tagged with 6FAM (OomF DavDC6 – GAGGGACTTTtGGGTAATCA) with OomR for about 1600 bp TRFLP amplicons and then use the below primers for nested detection of particular species.

(Actually ordered Pythium irregulare: PythIrreg Fwd – CGGAGCGGCGGGTGCTGTK with PythIrreg Rev – CaCCGAagtcgCcgACAG) [adapted from, “Species-specific PCR primers for Pythium developed from ribosomal ITS1 region”]

(Actually ordered Pythium ultimum: PythUltim Fwd – ACGAAGGTTGGTCTGTTG with PythUltim Rev – TCTCTACGCAACTAAATGC) [adapted from, “DETECTION OF PYTHIUM ULTIMUM FROM SOIL USING PCR MICROPLATE HYBRIDIZATION WITH SPECIES-SPECIFIC PRIMERS”]

(Actually ordered Phytophthora fragariae: PhytopFrag Fwd– with cACTTAGTTGGGGGCCTGTC with PhytopFrag Rev – CGCCGACTGGCCACACAG) [adapted from, “Validation and Exploitation of a PCR-based Diagnostic for Detecting Phytophthora in Strawberry and Raspberry”]

(Actually ordered Phytophthora infestans: PhytopInfest Fwd – TGGGCGAGCCCTATCAAAA with PhytopInfest Rev – CGATTCAAATGCCAAGCTA)[adapted from, “Development of a species-specific and sensitive detectionassay for Phytophthora infestans and its application for monitoring of inoculum in tubers and soil”]

Also gonna order fresh 27F Degen with a 6FAM label on it: AGRRTTYGATYHTGGYTYAG (noted that I put a typo into the sequence I used in my PhD)



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