Tissue genomic dna extraction kit

Genomic DNA was extracted from PDX tissues for STR analysis with a tissue genomic DNA extraction kit from Tiangen Biochemical Technology Co., Ltd. (Beijing, China). To evaluate gene expression, total RNA isolated from the PDX tissues was used for RT-PCR as previously described [3 (link), 23 (link)]. The primer sequences of the target genes (AR, AR7, JARID1D) are listed in Supplementary Table 1.

Genomic DNA was extracted from muscle tissue dissected from the body of a single adult W. pigra leech using a tissue genomic DNA Extraction Kit (TIANGEN, China). Samples of embryos at defined developmental phases were frozen with liquid nitrogen and RNAs were extract using RNAprep pure Tissue Kit (TIANGEN, China) in accordance with the standard protocol. The quality and quantity of DNA and RNA were tested by Nanodrop 2000 spectrophotometer (Thermo, USA) and 1% agarose gel electrophoresis. The Reverse transcription system Kit (Promega, USA) was used to synthesize the first strand of cDNA.
For genome sequencing, 260 bp paired-end shotgun libraries were prepared for sequencing on an Illumina NovaSeq 4000 platform to generate an initial survey based on a 19-K-mer distribution. The software Jellyfish (v2.1.4) [14 (link)] was used for counting k-mers, and the software GenomeScope (v1.0) [15 (link)] was used for estimating the genome size. Through whole-genome sequencing, a DNA library was constructed using the standard protocol of PacBio. RNA-seq libraries were generated using Vazyme kit and sequenced using NovaSeq 6000 platform (Illumina, USA). Sequencing data have been deposited in the NCBI Sequence Read Archive.

BPS (99% purity) was purchased from Sigma-Aldrich (Shanghai, China). Sesame oil (Jinlongyu, Shenzhen, China) was stored in glass bottles without bisphenol compounds. Phenylmethylsulfonyl fluoride (PMSF), 4% paraformaldehyde, 2.5% glutaraldehyde, BCA kit and anti-BDNF (Cat No: GB11559) antibody were purchased from Servicebio Technology (Wuhan, China). Anti-TrkB (Cat No: 13129-1-AP), anti-β-actin (Cat No: 20536-1-AP) and anti-CREB (Cat No: 12208-1-AP) antibodies were purchased from Proteintech (Chicago, IL, USA). Anti–phosphorylated (p)-CREB (Cat No: ab32096), anti-DNMT3a (Cat No: ab188470) and anti-DNMT3b (Cat No: ab2851) antibodies were purchased from Abcam (Shanghai, China). Anti-DNMT1 (Cat No: 5032) antibody was purchased from CST (Wuhan, China). The tissue genomic DNA extraction kit (Cat No: DP304) was purchased from Tiangen Biotech (Beijing, China). The other chemicals used in this study are commercially available at the required grade.