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#1
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| I'm trying to insert a fragment into an expression vector, the pET100 from invitrogens. then to ligation and transformation at the moment to analyze the colonies, these contain only the empty vector. The vector insert ratio was 1:1, 1:2, 2:1. The fragment to insert (~2.6KB) I get from PCR amplification using KOD Hot Start Polymerase. Someone can help me know what I'm doing wrong? Feel free to ask any information that I have forgotten and consider important. Thanks |
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#2
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| hi! OMG! What type of digests are you using? I mean blunt ended or the ones with overhangs? |
| Tags |
| cloning , expression , ligation or insertion , pet100 , problem , vector |
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