Keyword Analysis & Research: colony pcr streptomyces

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Frequently Asked Questions

How can I extract DNA from Streptomyces colonies for PCR?

For routine Streptomyces PCR, my lab does not extract DNA. It's laborious and time consuming, especially if you have many strains to check. Instead, try grinding actively growing colonies in DMSO, and proceed to use some of the resulting cell slurry as PCR template. This works quite well, is inexpensive, and saves time.

How do you screen for editing errors in Streptomyces clones?

Quick screening of the correctly edited clones can be achieved by following a Streptomyces colony PCR protocol. First, use a sterile wooden toothpick to scratch ~4 mm 2 of mycelia from the ISP2 plates from Step 33 (after incubation for 3–5 d) and transfer the mycelia to a PCR tube containing 20 µL DMSO.

Can CRISPR be used to manipulate Streptomycetes for bioactive natural products?

To develop advanced genetic manipulation methods for exploiting new bioactive natural products from streptomycetes, we first developed a toolkit using classic DNA double-strand break (DSB)-based CRISPR–Cas9 and its relatives 9.

Does DNA repair govern genome plasticity in Streptomyces?

Hoff, G., Bertrand, C., Piotrowski, E., Thibessard, A. & Leblond, P. Genome plasticity is governed by double strand break DNA repair in Streptomyces. Sci. Rep. 8, 5272 (2018).

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