{GENE PREP}

I.       Purpose:  To extract genomic DNA from plant material.

II.    Chemical Principal:  Modified CTAB method.

IIIA.  Preparation of Tissue Digest (off-line CTAB digestion)

       1. Put plant material (50-300mg(wet)*/hole) into AutoGen's tube unit, freeze in liquid nitrogen and
   grind the frozen samples in AutoGen's automated sample grinder or using    a set of pestle and mortal.

        2. Add 0.45ml of PL-M1 (Plant Lysis Solution) to the ground samples and digest at 65°C for
   30-60 minutes in a shaking incubator.

IIIB.  Preparation of Tissue Digest (on-board CTAB digestion)

       1. Put plant material (50-300mg(wet)*/hole) into AutoGen's tube unit, freeze in liquid nitrogen and
   grind the frozen samples in AutoGen's automated sample grinder or using a set of pestle and mortal.

        2. Load the tube unit onto the GENE PREP.

IV. Protocol Parameters:

A.    Sample Volume: 0.5 ml of CTAB/digest.

B.    Maximum Number of Samples: 48 samples per run
                                                     192 samples per run (with an optional Tube Unit Stacker)

C.    Processing Time:     2 hours for 48 samples (including 60 minute drying period).
                                  8 hours for 192 samples (including 60 minute drying period).

D.    Yield:     10-20 mg genomic DNA / 100 mg of young rice plant.
              10-20 mg genomic DNA / 300 mg Arabidopis thaliana.

E.     Quality:     OD_260/280 values are 1.7-1.9.
                  The DNA can be used directly in downstream processes such as fluorescence
                  DNA sequencing, PCR and restriction enzyme digestions*.
                  * The DNA from poly-saccharide rich samples might not be suitable for some downstream
                     applications.  Version 2 protocol is available for these type of plant materials.

V.  Running the Protocol:

        A.    Load Reagents and Samples
        B.    Select a Protocol
        C.    Enter Number of Samples
        D.     Start the Run

VI. Example of extracted genomic DNA

VII. Extraction and Purification Process: