Total RNA was extracted by TRIzol Isolation Reagent (Invitrogen). First-strand complementary DNA from total RNA was synthesized using Superscript III reverse transcriptase (Invitrogen) and Oligo-dT17 as the primer. Lipid metabolism gene expression was determined using the Mouse Fatty Acid Metabolism RT2 Profiler PCR Array (Qiagen). Primers used for real-time PCR were as follows: BDNF, 5′-ATGTCTATGAGGGTTCGGCG-3′ (forward) and 5′-GCGAGTTCCAGTGCCTTTG-3 (reverse); actin, 5′-CTGTCGAGTCGCGTCCA-3′ (forward) and 5′-ACCCATTCCCACCATCACAC-3′ (reverse); β-actin, 5′-AACCGTGAAAAGATGACCCAGAT-3′ (forward) and 5′-CACAGCCTGGATGGCTACGT-3′ (reverse). Real-time PCR was performed using 5 PRIME RealMasterMix SYBR ROX (Thermo Fisher Scientific) on an ABI7500 Real-time PCR System (Applied Biosystems, Foster City, CA).

mRNA sequencing was performed using the Illumina NextSeq v2 High SR75 (Illumina, San Diego, CA). Adapters were trimmed, and low-quality bases were removed from raw reads using Trimmomatic. Processed reads were then mapped to the mouse reference genome (National Center for Biotechnology Information build 37.2) using TopHat and Cufflinks. Differential gene expression was evaluated using Cuffdiff.

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.