Restriction Digest: Partial

Abstract
This protocol can be used to generate genomic DNA fragments of different sizes using a common cutter. This is useful for many screening/probing processes.

Materials

 * 10 microcentrifuge tubes

Reagents

 * Restriction Endonuclease
 * 10X Restriction Endonuclease buffer
 * BSA (10 mg/mL)
 * Genomic DNA (Concentration > 0.1 μg/μL)

Procedure

 * 1) Label the microcentrifuge tubes 1 - 10
 * 2) Mix together the following and invert to mix:
 * 3) 180μL Genomic DNA
 * 4) 20μL 10X Buffer
 * 5) 2μL BSA
 * 6) Dispense 40μL of this mix into tube 1 and 20μL into tubes 2 - 10 **[[Image:Critical step.png]]From this point keep everything on ice!!!
 * 7) Add 1μL of restriction-endonuclease to tube 1 and invert to mix.
 * 8) Transfer 20μL from tube 1 to tube 2 and invert to mix.
 * 9) Repeat the last step transfering 20μL from the preceding tube to the succeeding tube mixing with every transfer.
 * 10) Finally remove 20μL from tube 10
 * 11) Incubate all 10 tubes for 1 hour at 37°C.
 * 12) Finish the digest by submerging all 10 tubes in an 70°C water bath for 20 minutes.
 * 13) Run the samples on a gel and choose the size which best fits your application.

Acknowledgments
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Specific Protocols
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