Quick-DNA™ Fungal/Bacterial Miniprep Kit

Protocol tips

Upstream tips	Protocol tips	Downstream tips
		- Include RNAse treatment for 15-20 min. - Use prewarmed TE buffer to elute the DNA

Downstream tips
- Include RNAse treatment for 15-20 min. - Use prewarmed TE buffer to elute the DNA

Publication protocol

Isolation and purification of S. boulardii genomic DNA
A sachet of Dr. Reddy’s Laboratories Econorm 250 mg (B. No. 1500, Mfg. Date: 05/12, Expiry Date: 04/14) containing lyophilized cells of Sb was used as the source of the probiotic yeast. Yeast cells were suspended in Milli-Q water, serially diluted, and plated on Yeast Mold (YM) agar (Difco) plates. The plates were incubated at 37°C for 48 hours. An isolated colony was picked from the plate and cultured in Yeast Extract-Peptone-Dextrose (YEPD) broth (HIMEDIA) for 24 hours at 37°C in a rotary shaker (180 RPM). The cells were centrifuged at 5000 g for 10 minutes and washed with distilled water. DNA isolation was performed using the ZR Fungal/Bacterial DNA mini prep kit (Zymogen) as per instructions in its user manual. After isolation, the genomic DNA was treated with RNase A (1 μl of a 10 μg/mL stock solution for 100 μl of solution containing DNA) and incubated at 37°C for 30 minutes. Then, 1/10 volume of 3 M sodium acetate (pH 5.2) and 2.5 volumes of absolute ethanol was added followed by incubation at −20°C overnight and centrifugation at 14,000 rpm for 30 minutes at 4°C. Supernatant was carefully discarded; pellet was rinsed with 70% ethanol and centrifuged again at 14,000 rpm for 15 minutes at 4°C. The ratio of OD at 260/280 nm was ~1.8 as observed by NanoDropND-1000 spectrophotometer.

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Manufacturer protocol

Download the product protocol from Zymo Research for Quick-DNA™ Fungal/Bacterial Miniprep Kit below.

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