Single Oocyte Bisulfite Mutagenesis

Single oocyte bisulfite mutagenesis.

Epigenetics encompasses all heritable and reversible modifications to chromatin that alter gene accessibility, and thus are the primary mechanisms for regulating gene transcription. DNA methylation is an epigenetic modification that acts predominantly as a repressive mark. Through the covalent addition of a methyl group onto cytosines in CpG dinucleotides, it can recruit additional repressive p...

[31] Localized Mutagenesis 457 [31] Directed Mutagenesis with Sodium Bisulfite

With the development of recombinant DNA technology and the many analytic techniques based on restriction endonucleases, it has become relatively routine to isolate a segment of genomic DNA carrying a gene of interest, to define the limits of the gene within this DNA segment, and then to determine the gene's complete nucleotide sequence. In this way, a number of structural questions can be direc...

Astonishing fertility from a single oocyte recovery.

A 36 year old patient underwent in-vitro fertilization (IVF) and embryo transfer. Three embryos were transferred to her uterus and eight spare embryos were cryopreserved. The patient conceived a quadruplet pregnancy and delivered two boys and two identical twin girls by Caesarean section at 30 weeks gestation. The couple subsequently donated their cryopreserved embryos to the embryo donation pr...

Whole-Genome Bisulfite Sequencing

Next-generation sequencing technology has enabled genome-wide analysis of 5mC nucleotides at single-nucleotide resolution. Previous methods for whole-genome bisulfite sequencing (WGBS) yielded reduced genomic representation due to required DNA shearing, ligation of methylated sequencing adapters, and bisulfite conversion of unmethylated cytidine residues prior to sequencing. One disadvantage of...

BAT: Bisulfite Analysis Toolkit