Lipofectamine® 2000 Transfection Reagent

Protocol tips

Upstream tips	Protocol tips	Downstream tips
	Do not add antibiotics to the Transfection Medium

Protocol tips
Do not add antibiotics to the Transfection Medium

Publication protocol

2.7. Construction and transfection of the cytoglobin plasmid into rat mesangial cells
We used 2 primers (5′-TCCACCGGTATGGAGAAAGTGCCGGG-3′ and 5′-CATACCGGTCGGCCCTGAAGAGGGCAGA-3′) to obtain the gene of cytoglobin from the cDNA library of adult rat brain. The gene was then integrated in the vector (pCMV-2A-EGFP), and the sequence was further validated by sequencing. Then, the plasmid was transfected into rat mesangial cells (purchased from China Center for Type Culture Collection) via Lipofectamine 2000 as instructed. The transfected rat mesangial cells (obtained from ATCC) were cultured in the DMEM medium with 10% fetal bovine serum (Fig. ​(Fig.11B).

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