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脊髓挫伤损伤模型（小鼠，文献参考）

小鼠创伤性脊髓挫伤模型用于理解病理生理与测试干预；挫伤后小胶质细胞活化、病灶限制与胶质瘢痕反应为核心病理过程（PMID:40702215）。

Study design

以预先生成的 cDNA 为输入的 SYBR Green 实时定量 PCR 检测。每孔加入 10 ng cDNA，技术三重复；使用 PowerUp SYBR Green Master Mix 与 5 µM 正反向引物；按源方案循环条件扩增并读取 CT，以看家基因和参考实验组进行 ΔΔCT 分析。具体基因、引物和生物学分组按各研究设计确定。

Outcome measures

各样品目标序列的 CT 值；以看家基因归一化并以参考实验组计算 ΔΔCT/相对表达量。本方案不包含具体基因或疾病特异结论，输出判读依赖所选的引物、看家基因和实验分组。

Experimental animals

Experimental procedures

qPCR 反应体系 **Option 2****: SYBR qPCR** qPCR 反应体系 Prepare SYBR Master mix (ul per well): 1. 5ul PowerUp SYBR Green Master Mix (#A25776, Applied Biosystem-ThermoFisher) 1. 1ul Primers (Forward and Reverse, diluted in Nuclease-free water at 5µM) qPCR 反应体系 Add 6ul of SYBR Master mix to each well with an automatic pipette (Multipette E3, Eppendorf) qPCR 反应体系 Add 4ul of cDNA per well (10ng cDNA) in technical triplicates to each well of LightCycler 480 Multiwell 384-plate (#4729749001, Roche Diagnostics) 热循环 Perform PCR using the following cycling conditions in a LightCycler® 480 System (Roche): 1. 50˚C for 2min 1. 95˚C for 2min 1. 95˚C for 15s + 60˚C for 1min (x40cycles) 数据分析 Using the LightCycler Software obtain the threshold cycles (CT) signals from each samples. 数据分析 Perform ΔΔCT-method to analyze the expression data using the endogenous control genes and the reference experimental group.

Model identity

Model name
脊髓挫伤损伤模型（小鼠，文献参考）
Species
mouse

Biological characteristics

Research questions
脊髓损伤后炎症-修复反应与轴突通路重建机制；神经保护与再生干预评价。
Limitations
文献参考损伤范式，不代表公司实际保有、供应或质检；不记录致伤装置与操作细节。

Experimental methods

Experimental protocols

  • cDNA 模板的实时定量 PCR（qPCR，SYBR Green 分支）

    This protocol adopts the SYBR Green branch of the protocols.io qPCR v1 record. The input is pre-generated cDNA (10 ng per well); RNA extraction and reverse transcription are not included. PowerUp SYBR Green Master Mix and forward/reverse primers are used on a LightCycler 480 with 50˚C for 2 min, 95˚C for 2 min, and 40 cycles of 95˚C for 15 s plus 60˚C for 1 min. CT values are read and analysed by the ΔΔCT method. No specific genes, primer sequences or disease-model readouts are asserted.

Reagents and materials

References

  1. PMID:40702215; DOI:10.1038/s41598-025-12372-1; Sci Rep 2025
  2. Miquel Vila. Gene expression analysis by quantitative Real-Time PCR (qPCR). protocols.io. https://doi.org/10.17504/protocols.io.ewov1oyz2lr2/v1
  3. Bustin SA, et al. MIQE précis: Practical implementation of minimum standard guidelines for quantitative real-time PCR experiments. BMC Molecular Biology 2010;11:74. PMID 20858237. doi:10.1186/1471-2199-11-74
  4. ARRIVE 2.0 guidelines
Model consultation