Venous blood was collected and extracted for genomic DNA using a commercially available genomic DNA kit (Omega Bio-tek, Norcross, GA, USA). Six SNPs (rs1261085, rs1261084, rs8766, rs9960767, rs2958182, and rs12966547) of the TCF4 gene were detected based on their likely involvement in SCZ [2 (link), 18 (link), 19 (link)]. As previously described, genotyping was performed by SNPscan Genotyping Assays on an ABI 7900HT Fast Real-Time PCR System (Genesky Biotechnologies Inc., Shanghai, China) according to the manufacturer's instructions [20 (link)]. Polymerase chain reaction (PCR) was used to amplify the gene fragment containing the SNP loci of interest, followed by SNPscan of genotyping frequencies. We randomly chose 5% of samples with high DNA quality and repeated the analyses to guarantee the genotyping quality. The average genotype call rate for all markers was 97.2%.
Genomic dna kit
Manufactured by Omega Bio-Tek
Sourced in United States
The Genomic DNA Kit from Omega Bio-Tek is a laboratory product designed for the extraction and purification of genomic DNA from a variety of sample types. The kit provides a reliable and efficient method for obtaining high-quality genomic DNA for downstream applications such as PCR, sequencing, and other molecular biology analyses.
Automatically generated - may contain errors
Lab products found in correlation
Novaseq platform, by Illumina (2 mentions)
Nanodrop 2000 spectrophotometer, by Thermo Fisher Scientific (2 mentions)
Kod sybr qpcr mix kit, by Toyobo (1 mentions)
Trizol reagent, by Thermo Fisher Scientific (1 mentions)
Lightcycler 480 real time pcr system, by Roche (1 mentions)
Cytoplasmic nuclear rna purification kit, by Norgen Biotek (1 mentions)
Revertra ace qpcr rt master mix with gdna remover, by Toyobo (1 mentions)
Miseq machine, by Illumina (1 mentions)
Guaiacol, by Merck Group (1 mentions)
2 6 dmp, by Merck Group (1 mentions)
L dopamine, by Merck Group (1 mentions)
2 2 azinobis 3 ethylbenzothiazoline 6 sulfonic acid (abts), by Merck Group (1 mentions)
Gel extraction kit, by Omega Bio-Tek (1 mentions)
Miseq platform, by Illumina (1 mentions)
7 protocols using genomic dna kit
1
Genetic Analysis of TCF4 in SCZ
2
Comprehensive RNA and DNA Extraction Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The total RNA was extracted from tissue and cell samples using TRIzol® Reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer's instructions. Nuclear and cytoplasmic RNA were isolated from cells using a Cytoplasmic & Nuclear RNA Purification Kit (Norgen Biotek, Thorold, ON, Canada; Product # 21000, 37400). The concentration and purity of all RNA samples were subsequently measured using a NanoDrop 2000 spectrophotometer (Thermo Scientific, Wilmington, DE, USA). The corresponding cDNAs were generated using ReverTra Ace qPCR RT Master Mix with gDNA Remover (TOYOBO, Osaka, Japan). gDNA was extracted from cells using a Genomic DNA Kit (Omega Bio-Tek, Guangzhou, China) following the manufacturer's instructions. qRT‒PCR was performed on a LightCycler 480 Real-Time PCR System (Roche, Basel, Switzerland) using a KOD SYBR® qPCR Mix Kit (TOYOBO, Osaka, Japan) according to the manufacturer's protocol. The relative quantification of circRNA and messenger RNA (mRNA) expression was carried out by the 2-ΔΔCt method, and GAPDH was used as an internal control. All primer pairs were designed and synthesized by Tsingke Biotechnology Co., Ltd. (Beijing, China), and the sequences of the forward and reverse primers are listed in Supplemental Table 1
3
Gut Microbiome Analysis of Rat Feces
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
DNA from rat feces was extracted using a Genomic DNA Kit (Omega Bio-tek, GA, USA.). The primer pair (341 F, 5′-CCTAYGGGRBGCASCAG-3′; 806R, 5′-GGACTACHVGGGTWTCTAAT-3′) was used to amplify the 16S rRNA sequencing genes (V3–V4 regions) from the whole genome of bacteria. Quantification, pooling, and sequencing of amplicons were carried out on an Illumina MiSeq machine. FLASH (v1.2.8) program was used to stitch the double-ended sequences, and Vsearch (v2.3.4) filter was applied to eliminate unqualified sequences. Sequences with 97 % similarity are classified as OTUs. Based on systemic affinities of ITS2 gene sequence of RDP and Unite database, the 16S rRNA gene sequences were distributed into distinct taxonomic categories. PICRUSt software and KEGG database (https://huttenhower.sph.harvard.edu/galaxy/ ) were used to predict and analyze gene functions.
4
Cloning and Characterization of K. huakuii Enzymes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Strain K. huakuii LAM0618T was provided by the Agricultural Culture Collection of China (ACCC 06121). E. coli strains DH5α and BL21 (DE3) (Tian Gen Co., Ltd, China) were used as host strains for plasmid propagation and protein expression, respectively. The E. coli strains were routinely grown in Luria-Bertani medium. Antibiotics were added at desired concentrations (50 μg/mL kanamycin or 100 μg/mL ampicillin). A genomic DNA kit, a gel extraction kit and a plasmid kit were purchased from Omega Bio-Tek (Norcross, GA, USA). Enzymes for cloning procedures and protein ladders were obtained from Fermentas (St. Leon-Rot, Germany). ABTS, SGZ, 2,6-DMP, guaiacol and l -dopamine were purchased from Sigma-Aldrich (St. Louis, MO, USA). Other chemicals were of analytical grade or higher and were purchased from commercial sources.
5
Gut Microbiome Analysis of Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The 5 mice were selected from Ex and MCP-M groups at random, and DNA from feces was extracted by using a Genomic DNA Kit (Omega Bio-tek, Inc., Norcross, GA, USA). The 16S rRNA genes (V3-V4 regions) were amplified from the whole genome via the primer pair (341 F, 5′- CCTAYGGGRBGCASCAG-3′; 806R, 5′-GGACTACHVGGGTWTCTAAT-3′). All the amplicons were purified, quantified, and sequenced on an Illumina novaseq platform (San Diego, CA, USA). The barcode and connector sequence were removed. FLASH (v1.2.8) was used to stitch the double-ended sequences, and Vsearch (v2.3.4) was used to filter out the unqualified sequences (8 (link)). Finally, the sequence with 97% similarity was classified as an OTU. 16S rRNA gene reads were down-sampled to a read depth of 44,367 reads/sample and reads mapped to 16S OTUs (1,635 reads), to ensure sample compatibility regardless of sampling depth (21 (link)). Gene functions were predicted by PICRUSt.2 and analyzed by KEGG pathway enrichment analysis.1
6
Microbiome Profiling from Mouse Feces
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The other 5 mice fecal samples were collected and DNA from the mice feces was extracted by using a Genomic DNA Kit (Omega Bio-tek, Inc., Norcross, GA, USA), as previously stated [16 (link),17 (link)]. The 16S rRNA genes (V3-V4 regions) were amplified from the whole genome via the primer pair (341 F, 5′- CCTAYGGGRBGCASCAG-3′; 806R, 5′-GGACTACHVGGGTWTCTAAT-3′) with the barcode. All the amplicons were purified, quantified, and then sequenced on an Illumina novaseq platform and 250 bp paired-end reads were generated. The barcode and connector sequence were cleared away, and the double-ended sequences were stitched by using FLASH (v1.2.8) and the unqualified sequences were filtered out. Finally, the sequence with 97% similarity was classified as an OTU, and the systemic affinities of the ITS2 gene sequence of RDP and Unite database were used for distributing the 16S rRNA genes into distinct taxonomic categories.
7
Gut Microbiome Analysis in Rats
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The intestinal contents of five rats were collected, and the Genomic DNA Kit (Omega Bio-tek, Inc., Norcross, GA, United States) was used to extract DNA from the rats’ feces, as previously mentioned (Fulle et al., 2000 (link); Liu et al., 2021 (link)). The Primers were then used to amplify the 16S rRNA gene (V3-V4 region) from the whole genome (F,5-ACTCCTACGGGAGGCAGCA-3R,5-GGACTACHVGGGTWTCTAAT-3). After fluorescence quantification, all amplicons were purified, recovered, and sequenced on the Illumina MiSeq platform.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!