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primers fpr sequencing


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4 replies to this topic

#1 saad

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Posted 30 March 2009 - 02:54 PM

Hi everyone
I had have inserts in some vector like pMW172, pcDNA3 and pcDNA3 FLAG, and I want before starting gene expression to check the inserts by sending them for sequencing so I need a help to find forward and reverse primers for these vector/ inserts.

Thank you in advance

#2 bob1

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Posted 30 March 2009 - 04:09 PM

look at the plasmid map, look for names like M13 forward or sp6/t7 or bovine growth hormone A (BGHA)

#3 jiajia1987

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Posted 30 March 2009 - 05:01 PM

you can even design a primer of your own for sequencing based on the map!

#4 molgen

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Posted 31 March 2009 - 06:08 AM

pcDNA has a T7 sequencing primer.
If I remember correctly, pMW172's back bone is pET. If I'm right, then it to has a T7 sequencing primer.

#5 Nrelo

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Posted 03 April 2009 - 03:13 AM

it also depends on the size of your insert, if it is a bit large, say >1.5kb, you better design two more (one forward, one reverse, 18~21bp each) which will anneal to your insert.




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