Sequencing pDNA - (Jun/27/2008 )
Hi Everyone,
I am beginner in cloning experiments, and very excited to see the desired restriction map for the oligo (shRNA) cloned in to my vector, so would like to get it sequenced. And there are a few prior questions -
1) Would I have to linearise the plasmid and then send it for sequencing. (I mean which is the right way)
2) And what quantity of pDNA should be submitted for such an analysis.
I am sure yours inputs will make me learn things.
Thanks in advance
I am beginner in cloning experiments, and very excited to see the desired restriction map for the oligo (shRNA) cloned in to my vector, so would like to get it sequenced. And there are a few prior questions -
1) Would I have to linearise the plasmid and then send it for sequencing. (I mean which is the right way)
2) And what quantity of pDNA should be submitted for such an analysis.
I am sure yours inputs will make me learn things.
Thanks in advance
hi there
1) no need to linearise
2) I use about 300-400ng - the place where you are sending your sequencing should provide you with all the details about quantity etc.
Good luck
Clare
1) no need to linearise
2) I use about 300-400ng - the place where you are sending your sequencing should provide you with all the details about quantity etc.
Good luck
Clare
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Thank you Clare,
The sequencing laboratory is in the other part of city, I will follow your instructions for sure.
Thank you Clare,
The sequencing laboratory is in the other part of city, I will follow your instructions for sure.
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Hope it all works out 