Comprehensive Gene Expression Analysis

Complementary DNA (cDNA) was synthesized by a Verso cDNA RT kit (Cat# AB1453B, Thermo Scientific) per the manufacturer's protocol. Real-time PCR for target mRNAs was performed using TaqMan gene expression assays for Lipocalin 2 (Lcn2, Mm01324470_m1), Ccl2 (Mm00441242_m1), Ccl7 (Mm00443113_m1), Gbp2 (Mm00494576_g1), IL-1β (Mm0133-6189_m1), Myc (Mm00487804_m1), IL-6 (Mm00446190_m1), suppressor of cytokine signalling 3 (SOCS3, Mm00545913_s1), C5ar1 (Mm00500292_s1), Trem2 (Mm04209424_g1), Ptgs2 (Mm00478374_m1), Nlrp3 (Mm00840904_m1), Mb21d1 (Mm01147496_m1), Tmem173 (Mm00727224_s), Tnfrsf1 (Mm00441883_g1), Fos (Mm00487425_m1), Tgm2 (Mm00436979_m1), Lamp1 (Mm01217070_m1), Pink1 (Mm00550827_m1), Apoe (Mm01307192_m1), Blnk (Mm01197846_m1), and GAPDH (Mm99999915_g1) (Applied Biosystems, Carlsbad, CA] on an QuantStudio™ 5 Real-Time PCR System (Applied Biosystems). Gene expression was normalized by GAPDH and compared to the control sample to determine relative expression values by the 2-ΔΔCt method 31 (link), 41 (link).

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Li Y., Lei Z., Ritzel R.M., He J., Li H., Choi H.M., Lipinski M.M, & Wu J. (2022). Impairment of autophagy after spinal cord injury potentiates neuroinflammation and motor function deficit in mice. Theranostics, 12(12), 5364-5388.

Publication 2022

Verso cDNA RT kit Pink1 GAPDH QuantStudio™ 5 Real-Time PCR System

Apoe Assays Blnk C5ar1 Ccl2 Ccl7 Cdna Cytokine Gapdh Gene expression Il 1β Lamp1 Lipocalin 2 Mb21d1 Mrnas Ptgs2 Real time pcr Tgm2 Trem2

Corresponding Organization :

Other organizations : University of Maryland, Baltimore

Top 5 similar protocols

Variable analysis

independent variables

Tissue samples/experimental conditions

dependent variables

Expression levels of the following target mRNAs:
Lipocalin 2 (Lcn2)
IL-1β
SOCS3
C5ar1
Trem2
Ptgs2
Nlrp3
Mb21d1
Tmem173
Tnfrsf1
Lamp1
Pink1

control variables

GAPDH expression (used for normalization)

controls

Positive control: Not specified
Negative control: Not specified

Annotations

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