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Old 06-05-2019, 08:22 AM   #1
dacotahm
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Location: ND, USA

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Red face Alternative to Qiagen Puregene Tissue kit for HMW gDNA?

Hello,

We're extracting high molecular weight gDNA for PacBio from an insect-a bee- using the Qiagen Puregene Tissue kit (4g). We have trouble with this kit back-ordered all the time. Could someone from the community recommend an alternative for HMW gDNA that works well on tissue for PacBio?

Thanks,

Dacotah
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Old 06-05-2019, 08:34 AM   #2
ATϟGC
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I have tried the following protocol with insects and it works quite well in terms of purity and length:

Modified low‐salt CTAB extraction of high‐quality DNA from contaminant‐rich tissues

https://onlinelibrary.wiley.com/doi/...755-0998.12616

A benchtop protocol is available in the "Supporting Information" section.
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Old 06-06-2019, 04:28 AM   #3
olafblue1955
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Hi Dacotah,

Qiagen's PureGene kits are nearly identical to the former Epicentre MasterPue DNA Purification kits, which are now available through Lucigen/Biosearch Technologies. Catalog numbers would be MC85200 for 200 reactions and MC89010 for 10 reactions. Both are non-column based products.

Olafblue1955
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Old 06-06-2019, 09:09 AM   #4
dacotahm
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Oh, that's great, thank you very much.
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extraction, gdna, hmw, pacbio, qiagen

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