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| Recently we have been having an issue with XL10-Gold. The problem is that once you spin them down, they don't seem to want to be resuspended. Anyone else noticed that? Probably wounldn't be much of an issue--but our preps are done in 384-well deep plates. I always thought the "XL" host strains were great sequencing strains. But now I think that may be entirely based on my experience with XL1. I don't recall using XL2. The transition from XL1->XL2->XL10 seems largely to involve the addition/mutation of genes to increase chemical transformation efficiency. According to patent descriptions on the web, the "Amy" gene codes for amylase and when transfered to a host strain, increases its transformation efficiency several-fold. "Hte" also increases transformation efficiency. Any reason why an amylase gene would make pellets sticky? Any advice on decreasing the stickness of the pellets? Here are the genotypes: XL1-Blue MR Genotype: ?(mcrA)183 ?(mcrCB-hsdSMR-mrr)173 endA1 supE44 thi-1 recA1 gyrA96 relA1 lac XL1-Blue MRF´ Kan Genotype: ?(mcrA)183 ?(mcrCB-hsdSMR-mrr)173 endA1 supE44 thi-1 recA1 gyrA96 relA1 lac [F´ proAB lacIqZ?M15 Tn5 (Kanr)] XL2-Blue MRF´ Genotype: ?(mcrA)183 ?(mcrCB-hsdSMR-mrr)173 endA1 supE44 thi-1 recA1 gyrA96 relA1 lac [F´ proAB lacIqZ?M15 Tn10 (Tetr) Amy Cam-r] XL10-Gold Kan-r Genotype: Tetr D(mcrA)183 D(mcrCB-hsdSMR-mrr)173 endA1 supE44 thi-1 recA1 gyrA96 relA1 lac Hte [F’ proAB lacIqZDM15 Tn10 (Tetr) Tn5 Kan-r Amy] -- Phillip |
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| bacteria , give , pellets , sticky , xl10gold |
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