Molecular Biotechnology Workshop

When
July 9-13

Where
College of Veterinary Medicine, Veterinary Science Building

Fee
$1125 through July 1, $1225 after July 1
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Lecture Schedule: 8:00  - 10:30 a.m.

Date              Topic

July 9               Organization and structure of biological information
July 10              DNA synthesis, replication and PCR
July 11              Recombinant DNA basics – from life to lab
July 12              Gene expression
July 13              Real-time PCR, DNA microarrays, and Bioinformatic analyses     

Laboratory Schedule: 10:30 a.m. - 5:00 p.m.

Date                 Topic
July 9                 Good lab practice.  Isolation and analysis of DNA.
July 10               DNA synthesis by PCR.  Restriction digest of DNA.
July 11               DNA sequencing. Recombining DNA.
July 12               RNA purification. Bacterial transformation. Real-time PCR.
July 13               Bioinformatics. Genetic analysis.

Monday Laboratory
Lab 1A: Plasmid Extraction (Animal)
        I: Extraction of plasmids from culture (Qiagen kit)
        II: Agarose gel to determine concentration

Lab 1B: Genomic DNA Extraction (Plant)
        I: Extraction of genomic DNA from powdered oat tissue
        II: Agarose gel to visualize difference between genomic DNA and plasmidI
        III: Set up PCR reaction   

Tuesday Laboratory
Lab 2A: Sequence sample preparation (Animal)
        I: Set-up of samples of plasmid for sequencing

Lab 2B: Genomic DNA extraction (Plant) for PCR (continued from lab 1Bb)
        I: Agarose gel of PCR

Lab 2C: Sensitivity of detection of transgenic material (Plant)
        I: Dilution of transgenic soybean with nontrangenic
        II: PCR set-up using primers specific to transgene bar (Qiagen Master Mix)

Lab 2D: Analysis of pGLO and pSK constructs (Bacteria)
        I: Restriction enzyme of pGLO and pSK constructs (EcoRI and XbaI)
        II: Agarose gel to visualize digestion of plasmids

Wednesday Laboratory
Lab 3A: Tour of sequencing facility AGAC - 10:30am to 11:15pm (Animal)

Lab 3B: Sensitivity issue association with detecting transgenic material (Plant)
        I: Agarose gel to visualize PCR from soybean samples

Lab 3C: Analysis of pGLO and pSK construct (Bacteria)
        I: Digestion clean up - Qiagen PCR Clean up kit
        II: Ligation of pGLO and pSK

Thursday Laboratory
Lab 4A: Expression analysis of transgene in soybean material (Plant)
        I: Extraction of total RNA from soybean material (Qiagen kit)
        II: Set-up of reverse-transcriptase PCR (Qiagen one-step)

Lab 4B: Transformation of pGLO, pSK and pGLO+pSK constructs (Bacteria)
        I: Chemical competent cells multiply above plasmids (Invitrogen TOP 10)

Lab 4C:  Real-time PCR

Friday Laboratory
 Lab 5A: Bioinformatics analysis of sequence material (Animal)
        I: Computer analysis

Lab 5B: Expression analysis of transgene in soybean material (Plant)
        I: Agarose gel of RT-PCR

Lab 5C: Analysis of pGLO, pSK, and pGLO+pSK constructs (Bacteria)

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