Total RNA from human endometrium and mouse uterine samples was extracted using the RNeasy Mini Kit (Qiagen Ltd) according to manufacturer’s instructions. RNA samples were reverse transcribed using iScript cDNA synthesis kit (Bio-Rad Laboratories Ltd) alongside control samples. mRNA transcripts were quantified relative to appropriate reference genes (Human: SDHA and ATP5B, Mouse: RLP13 and ACTB), as determined by geNorm assay (Primerdesign Ltd, Southampton, UK). Specific primers were designed using the universal probe library assay design centre and checked with BLAST (Table 4). Reactions were performed in triplicate alongside controls using ABI QuantStudio system under standard conditions with TaqPath ProAmp Master Mix (Life Technologies). Quantification was performed using the 2-ΔΔCt method with normalisation against a sample of liver or placental cDNA.

Details of PCR primers and universal probe library probe number.

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.