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Miniprep and sequencing


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4 replies to this topic

#1 rimal

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Posted 21 August 2010 - 10:11 AM

Hi all,

I need to sequence my plasmid, can I do that after miniprep or do I need to clean up the DNA??

#2 ranvi

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Posted 21 August 2010 - 12:57 PM

If you check the concentration using nanodrop you can see the value of A260/280 if its around 1.8 then usually the plasmid is good enough to sequence
if you clean up from miniprep the yield will be very low to send for sequencing , I normally send the plasmid for sequencing with out further purification it works fine

Edited by ranvi, 21 August 2010 - 12:58 PM.


#3 rimal

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Posted 21 August 2010 - 02:33 PM

If you check the concentration using nanodrop you can see the value of A260/280 if its around 1.8 then usually the plasmid is good enough to sequence
if you clean up from miniprep the yield will be very low to send for sequencing , I normally send the plasmid for sequencing with out further purification it works fine



Thanks very useful info.

#4 christy

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Posted 22 August 2010 - 04:41 PM

Hi all,

I need to sequence my plasmid, can I do that after miniprep or do I need to clean up the DNA??



while doing miniprep in the washing step with PE buffer (qiagen) wash twice first time 750 ul and again 500 ul that is enough for sequencing. I do this.

#5 microgirl

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Posted 23 August 2010 - 05:52 AM

Miniprep is cleaning up your plasmid. I always just go ahead and sequence and have never had a problem.




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