Restriction Digest: Partial

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==Abstract==
 
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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. 
 
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==Materials==
 
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* 10 microcentrifuge tubes
 
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===Reagents===
 
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* Restriction Endonuclease
 
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* 10X Restriction Endonuclease buffer
 
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* BSA (10 mg/mL)
 
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* Genomic DNA (Concentration > 0.1 μg/μL)
 
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==Procedure==
 
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# Label the microcentrifuge tubes 1 - 10
 
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# Mix together the following and invert to mix:
 
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## 180μL Genomic DNA
 
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## 20μL 10X Buffer
 
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## 2μL BSA
 
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# 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!!!'''
 
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# Add 1μL of restriction-endonuclease to tube 1 and invert to mix.
 
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# Transfer 20μL from tube 1 to tube 2 and invert to mix.
 
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# Repeat the last step transfering 20μL from the preceding tube to the succeeding tube mixing with every transfer.
 
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# Finally remove 20μL from tube 10
 
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# Incubate all 10 tubes for 1 hour at 37°C.
 
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# Finish the digest by submerging all 10 tubes in an 70°C water bath for 20 minutes.
 
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#Run the samples on a gel and choose the size which best fits your application.
 
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==Acknowledgments==
 
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Acnkowledge any help you had in development, testing, writing this protocol.
 
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==References==
 
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See [[OpenWetWare:Biblio]] for information on how to reference within a wiki.
 
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==Specific Protocols==
 
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Add links to all the OWW protocols that have been used in making the consensus.
 
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==Discussion==
 
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You can [[Talk:{{PAGENAME}}|discuss this protocol]].
 
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[[Category:Protocol]]
 
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[[Category:In vitro]]
 
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[[Category:DNA]]
 

Revision as of 07:15, 18 June 2009

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