Thirty-two gauge catheters were inserted through the atlanto-occipital membrane and extended to T8 (36 (link)). The location of each catheter was confirmed following euthanasia. Rats received either BDNF antagonist trkB-Fc (R&D Systems, Minneapolis, MN), 5 µg in 10 µl sterile saline or vehicle once/day during the 9-day stress period or BDNF siRNA or control siRNA (2 µg of the appropriate siRNA; 1:5 v/v with i-Fect transfection reagent Neuromics, Edina, MN). Each rat received 2ug siRNA/10 µl.) on days 1, 3, 5 and 7 of the 9 day stress period.
I fect transfection reagent
Manufactured by Neuromics
I-Fect is a transfection reagent designed for efficient delivery of nucleic acids, such as plasmid DNA or small interfering RNA (siRNA), into a variety of cell types. It facilitates the uptake of genetic material into cells, enabling gene expression studies or gene silencing experiments.
Automatically generated - may contain errors
Lab products found in correlation
2 protocols using i fect transfection reagent
1
Intraspinal Catheter Implantation and BDNF Modulation
2
Intrathecal miRNA Administration Protocol
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miRNA administration protocol was adapted from previous report of intrathecal miRNA delivery [27 (link)]. To administer miRNA mimics, a polyurethane catheter (25G, 5.5 cm long, SAI infusion) was placed into the intrathecal space of the lumber L4–L5 vertebrae under isoflurane anesthesia. The catheter was stereotactically secured under the skin and occluded between injections. A custom miRCURY (Exiqon) miR-126 mimic containing a 5′ cholesterol tag and 3′ fluorescein label was injected at 2 nmol concentration with 4 µl iFECT transfection reagent (Neuromics). A total of 6 µl was delivered into the catheter connection juncture using a 25G blunt end needle on a Hamilton syringe. The catheter was then flushed with 7 µl sterile PBS to ensure miRNA reached the intrathecal space.
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