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> sequencing primer: T7 or T7promoter?
kristian
post Nov 4 2009, 06:36 PM
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I am going to send my recombinant plasmid to a sequencing facility. I used pGEM Teasy vector which contains a T7 promoter. The form that I am filling up gives 2 options for T7 as sequencing primer namely T7 (AATACGACTCACTATAG) and T7 Promoter (TAATACGACTCACTATAGGG). These primers target the same site in the plasmid but vary a bit in length. Please help me choose which should I use.
Thanks!
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Quasimondo
post Nov 4 2009, 10:25 PM
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QUOTE (kristian @ Nov 5 2009, 11:36 AM) *
I am going to send my recombinant plasmid to a sequencing facility. I used pGEM Teasy vector which contains a T7 promoter. The form that I am filling up gives 2 options for T7 as sequencing primer namely T7 (AATACGACTCACTATAG) and T7 Promoter (TAATACGACTCACTATAGGG). These primers target the same site in the plasmid but vary a bit in length. Please help me choose which should I use.
Thanks!

The longer the complementary sequence, the lower the chance you got the contaminated result, I think. So the longer sequence would be my choice
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jiajia1987
post Nov 4 2009, 10:50 PM
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I would choose T7 promoter as well.
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kristian
post Nov 7 2009, 12:13 AM
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QUOTE (Quasimondo @ Nov 5 2009, 02:25 PM) *
QUOTE (kristian @ Nov 5 2009, 11:36 AM) *
I am going to send my recombinant plasmid to a sequencing facility. I used pGEM Teasy vector which contains a T7 promoter. The form that I am filling up gives 2 options for T7 as sequencing primer namely T7 (AATACGACTCACTATAG) and T7 Promoter (TAATACGACTCACTATAGGG). These primers target the same site in the plasmid but vary a bit in length. Please help me choose which should I use.
Thanks!

The longer the complementary sequence, the lower the chance you got the contaminated result, I think. So the longer sequence would be my choice

Thanks for your help! smile.gif
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kristian
post Nov 7 2009, 12:14 AM
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QUOTE (jiajia1987 @ Nov 5 2009, 02:50 PM) *
I would choose T7 promoter as well.

Thanks! smile.gif
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