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Storage life of plasmid DNA in water


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#1 science noob

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Posted 12 December 2013 - 10:19 PM

How long can I store plasmid DNA reconstituted in water for? I still have the mushed up blotting paper in the tube together with some DNAse free water.  Storage conditions: 4 degrees C (or should I store it at -20?)



#2 Ameya P

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Posted 13 December 2013 - 12:43 AM

If you plan to use it much later (say after a week or so), -20 is the way to go, otherwise, 4 degrees should be fine. 


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#3 science noob

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Posted 13 December 2013 - 04:16 AM

If you plan to use it much later (say after a week or so), -20 is the way to go, otherwise, 4 degrees should be fine. 

What would be the best way to store palsmid DNA? (A) In it's original state in a tube (since everytime you only need a few microlitres for transformation), (B) Transformed competent cells on agar plate, © Transformed competent cells in glycerol or (D) plasmid prep after transformation?



#4 phage434

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Posted 13 December 2013 - 04:21 AM

I'd recommend both C and D.

 

Water is actually a bad choice for storing DNA. Use TE instead -- the Tris buffers at basic pH for better DNA stability, and the EDTA removes magnesium, a required cofactor for DNAse enzymes.



#5 science noob

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Posted 13 December 2013 - 03:08 PM

I'd recommend both C and D.

 

Water is actually a bad choice for storing DNA. Use TE instead -- the Tris buffers at basic pH for better DNA stability, and the EDTA removes magnesium, a required cofactor for DNAse enzymes.

what composition of glycerol should i use? 50%? or 10%? in LB?



#6 phage434

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Posted 13 December 2013 - 06:16 PM

15% glycerol in an LB culture is a good start.






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