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XL10-Gold bacteria give sticky pellets

XL10-Gold bacteria give sticky pellets - Protocols and Methods Forum

XL10-Gold bacteria give sticky pellets - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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Old 10-02-2005, 03:44 PM
Phillip SanMiguel
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Default XL10-Gold bacteria give sticky pellets



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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