Bisulfite sequencing was performed using specific tissue samples and cells. The primers used for bisulfite sequencing of the KvDMR1 fragment are listed in Supplemental Table 3 . The amplified area contained 28 CpG sites. Amplified PCR products were purified using a gel extraction kit (Takara) and ligated into the pMD19-T plasmid using the TA-cloning system (Takara). The sequences were analyzed using 3730 DNA Analyzer polymers (Applied Biosystems, Carlsbad, CA).
3730 dna analyzer polymers
Manufactured by Thermo Fisher Scientific
Sourced in United States
The 3730 DNA Analyzer is a high-performance capillary electrophoresis instrument designed for DNA sequencing and fragment analysis applications. The instrument utilizes specialized polymer solutions to facilitate the separation and detection of DNA fragments during the analysis process.
Automatically generated - may contain errors
Lab products found in correlation
Pmd19 t vector system, by Takara Bio (3 mentions)
Epitect bisulfite kit, by Qiagen (3 mentions)
Gel extraction kit, by Takara Bio (1 mentions)
Pmd19 t vector, by Takara Bio (1 mentions)
Tianamp genomic dna kit, by Tiangen Biotech (1 mentions)
Ez dna methylation gold kit, by Zymo Research (1 mentions)
5 protocols using 3730 dna analyzer polymers
1
Bisulfite Sequencing of KvDMR1
2
Methylation analysis of BDNF, NGF, and FOS in PCOS placentae
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
A methylation assay was performed at the promoter region of BDNF, NGF, and FOS genes in the foetal side of placentae from PCOS and control women using the bisulphite genomic sequencing (BSP) method as previously described [47 (link)]. Total genomic DNA was extracted using the TIANamp Genomic DNA Kit (DP304-02, Tiangen Biotech, China). Bisulphite treatment was conducted using EZ DNA Methylation-Gold Kit (D5005, ZYMO Research, USA). The methylation status of cytosine phosphate guanine (CpG) sites located on gene promoters were analysed by cloning and sequencing bisulphite-treated DNA. The modified DNA was amplified by PCR, and the products were purified and cloned into a pMD 19-T vector (3271, Takara, Japan). Ten colonies of each subject were randomly selected for the plasmid DNA and sequenced with 3730 DNA Analyzer Polymers (3730S, Applied Biosystems, USA, RRID: SCR_018052).
3
Methylation Analysis of Pax8 in Mice
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Genomic DNA was extracted from thyroids of 3 and 8-week-old mice in the control and Ev100 groups. EpiTect bisulfite kit (Qiagen, Valencia, CA) was used to deaminate cytosine to uracil according to the manufacturer’s instructions, while 5-methyl-cytosine was protected from deamination. Nested PCR was used to increase the specificity of DNA amplication (Primers are listed in Supplemental Table 1 ). Methylation status of the Pax8 CpG island was determined by cloning and sequencing of bisulfite-treated DNA. The purified PCR products were cloned using the pMD19-T vector system (TaKaRa, Dalian, China). The sequence obtained by cloning was analyzed with 3730 DNA Analyzer polymers (Applied Biosystems, Carlsbad, CA).
4
Methylation Analysis of MCAT DMR
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Genomic DNA was extracted from lymphocytes of umbilical cord blood in control and GDM groups. Bisulfite was converted using the EpiTect bisulfite kit (Qiagen, Valencia, CA) to deaminate cytosine to uracil according to the manufacturer’s instructions; 5-methyl-cytosine was protected from deamination. Analysis of the methylation status of the MCAT DMR was determined by cloning and sequencing of bisulfite-treated DNA. The purified PCR products were cloned using the pMD19-T vector system (TaKaRa, Dalian, China). The sequence obtained by cloning was analyzed with 3730 DNA Analyzer polymers (Applied Biosystems, Carlsbad, CA).
5
DNA Methylation Analysis of Mouse Placenta
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Genomic DNA was extracted from placenta of D18.5 mice in the control, F1-GDM, and GDM♂-GDM♀ groups. Bisulfite was converted using the EpiTect bisulfite kit (Qiagen, Valencia, CA) according to the manufacturer's instructions to deaminate cytosine to uracil; 5-methyl-cytosine was protected from deamination. The methylation status of Dlk1 differentially methylated region (DMR), Gtl2-DMR, and intergenic differentially methylated region (IG-DMR) was determined by cloning and sequencing of bisulfite-treated DNA. The full list of primer sequences is shown in Supplementary Table 2 . The purified PCR products were cloned using the pMD 19-T vector system (Takara, Dalian, China). The cloned sequence was analyzed with 3730 DNA Analyzer Polymers (Applied Biosystems, Carlsbad, CA).
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!