QIAamp DNA Micro 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

Flow cytometry and nested PCR of AT-SVF cells
For flow cytometry of AT-SVF T cells, isolated AT-SVF cells were stained for CD3-Pacblue, CD4-PerCPCy5.5, CD45RO-FITC, and CD69-APC mabs (Biolegend or BD Biosciences), and analyzed with appropriate controls using a Gallios Flow Cytometer (Beckman-Coulter). For nested PCR's, DNA was first extracted from AT-SVF cells with QIAamp Microkit (Qiagen). Nested PCR reactions were performed by 2 rounds of 35-40 cycles using primers spanning HIV LTR-Gag (RU5+SK39 outer and US5+US3 inner primers) [16], and Envelope C2V3C3 (PCR5+R344 outer and EN1+EN4B inner primers) regions [17]. PCR products were gel-purified and sequenced by the BCM DNA Sequencing Core.

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Manufacturer protocol

Download the product protocol from Qiagen for QIAamp DNA Micro Kit below.

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