{AutoGenFlex 3000}

I.       Purpose:    To extract and purify genomic DNA from mouse tail and other types of animal tissues.

II.    Chemical Principal:  Proteinase K/phenol extraction method.

III. Pretreatment of Mouse Tail:

A.    Cut 5 - 10 mm of tail (about 10 - 30 mg).

B.    Place tail in 500µl of a buffered Proteinase K solution (250µl of AGF TD-M1 including 0.4mg/ml Proteinase K and 250µl of AGF TD-M2).

C.    Incubate at 37°C - 55°C for 6 hours to overnight.

IV. Protocol Parameters:

A.    Sample Volume: 0.5ml of digest solution.

B.    Maximum Number of Samples: 40 samples per run (AutoGenFlex 3000).

C.    Processing Time: 3.4 hours for 40 samples (Includes 1 hour drying time)

D.  Yield: 20 - 60 µg of purified DNA per 10 - 30 mg of mouse tail.

E.     Quality:    Typical OD_260/280 values are ³ 1.8
             The DNA is RNA free and can be used directly in downstream processes such as
                fluorescence DNA sequencing, PCR, Southern blotting and restriction endonuclease digestions.

IV.     Running the Protocol:

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

V. Example of genomic DNA obtained from mouse tail.

A 10 mm sample (about 30mg) was pre-treated with proteinase K and loaded onto the AutoGen system for purification.  Approximately 2 mg DNA per mg of tissue was obtained with a purity of OD_260/280 = 1.8 to 1.9.

VI.    Extraction and Purification Process: