DNeasy Blood & Tissue Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
		- Include RNAse treatment for 15-20 min. - Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C - Use prewarmed TE buffer to elute the DNA

Downstream tips
- Include RNAse treatment for 15-20 min. - Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C - Use prewarmed TE buffer to elute the DNA

Publication protocol

2.1. Gene cloning
Genomic DNA of S. sviceus ATCC 29083 was isolated from a 0.5 ml DSM 924 bacterial cell culture using the DNeasy Blood & Tissue Kit (Qiagen). The gene encoding the acetyltransferase (ZP_05014886), termed SsAT, was amplified using the polymerase chain reaction (PCR) from S. sviceus genomic DNA. The forward primer had the sequence 5′-CCAGG­GACCAGCAATGTACGGGGCACGCGACCG-3′ and the reverse primer had the sequence 5′-GAGGAGAAGGCG­CGTTATCACCGGCCTTCTTCTGCCGTG-3′. The amplification product was cleaned using a QIAquick Gel Extraction Kit (Qiagen) and was cloned following a ligation-independent cloning (LIC) method (Bonsor et al., 2006 ▶; Fogg & Wilkinson, 2008 ▶) into the vector pETYSBLIC-3C, which incorporates a 3C protease-cleavable N-terminal hexahistidine tag

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Manufacturer protocol

Download the product protocol from Qiagen for DNeasy Blood & Tissue Kit below.

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