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DNA was extracted using AccuPrep[R] Genomic DNA Extraction Kit (Bioneer, Korea) and treated with bisulfite using EZ DNA Methylation Gold' (Zymoresearch, Orange, CA, USA).
Primer sequences used in bisulfite pyrosequencing Gene name Accession Primer PCR primer sequence (Symbol) number (5'-3') Peroxisome NM_181024 Forward tgaggtttgtggtgatga- proliferator- ttattt activated receptor gamma Reverse aacacaatttccccaaccatta isoform 1 (PPARG1) Fatty acid BT10868 Forward tttaatttttttgttagga binding attgggttat protein 4 Reverse aaaaacatacaacctaaatc- (FABP4) ccttaca Gene name Sequencing primer Target region: Number (Symbol) (5'-3') relative to of CpG transcription sites start site checked Peroxisome aacccaaataaataa- +144 to +225 3 proliferator- aattct activated receptor gamma isoform 1 (PPARG1) Fatty acid gttaggaattgggtt- -9,664 to -9,469 5 binding atatagta protein 4 (FABP4) PCR, polymerase chain reaction.
In this study, the test was optimized for use in a clinical laboratory, including improvements to the magnetic particle-based DNA extraction and the use of ammonium bisulfite in the conversion reaction.
First a mix of 42 mmol 4-t-butyl-2-(N-piperidinomethyl)cyclohexane and 21 mmol sodium bisulfite was solved in 4 ml water at ambient temperature (25[degrees]C) to obtain the sulfonic.
To determine whether DNA methylation correlates with Svs4 transcription, we used bisulfite sequencing to examine the methylation status of these four CpGs in WT SVs at weeks 3, 5, and 10.
Diallyl amine (5), on treatment with paraformaldehyde and sodium bisulfite, afforded sodium diallylaminomethanesulfonate (6), which on treatment with HCl gave a mixture of diallylammoniomethanesulfonate (7) and its corresponding sulfonic acid 8 in excellent yield (Scheme 2).
5, which could possibly influence the results derived from techniques based on restriction enzymatic digestion and/or bisulfite modification, comparing two different methods.
Several other water treatment plants have switched from sulfur dioxide gas to liquid bleach and sodium bisulfite.
com)-- Today Bioo Scientific announced the launch of the NEXTflex[TM] Bisulfite-Seq Kit which is the first complete kit available designed to simplify library preparation of bisulfite treated DNA for down-stream high-throughput sequencing.
The isolated DNA was subjected to methylation specific PCR (MS-PCR) after bisulfite treatment to determine the patterns of methylation status.
Bisulfite modification of the digested tissue samples (18 [micro]L) was performed using the EZ DNA methylation kit (Zymo Research Co, Orange, California).
The Cupric Inhibitor contains Thiourea (76 g/L) and Sodium Bisulfite (10 g/L) in deionized water.
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