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Problem with pART27 vector [View Printable]
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Vinny
Group: Member
Posts: 4
Joined: Jan 10, 2007
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I have a problem with cloning pHannibal cassette in pART27....I cut phannibal cassette and open pART27 with NotI and seems to be OK....after i treat the pART27 with CIAP, extract all from gel...ligate and cloning the ligation in E.coli DH5alfa...after i select on antibiotic Kanamicin 50ug/ml...but not works....There's anyone that can help me????
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 Posted Jan 10, 2007, 19:39 PM |
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Tony Rook
Group: Moderators
Posts: 582
Joined: Nov 03, 2005
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Vinny:
In the past, I have had problems with my antibiotic concentration being too high during selection. You may want to try and select at a lower concentrations such as 25 ug/ml.
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Tony Rook
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| Posted Jan 11, 2007, 19:00 PM |
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Rock
Group: Member
Posts: 1
Joined: Sep 07, 2007
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| Vinny said: | | I have a problem with cloning pHannibal cassette in pART27....I cut phannibal cassette and open pART27 with NotI and seems to be OK....after i treat the pART27 with CIAP, extract all from gel...ligate and cloning the ligation in E.coli DH5alfa...after i select on antibiotic Kanamicin 50ug/ml...but not works....There's anyone that can help me???? |
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| Posted Sep 07, 2007, 11:20 AM |
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