08/31 Lab meeing-小嬿

1. to observe whether CTCF interact with EBV Rta by Immunoprecipitation assay

a) in TW01 inducible cell lines, CTCF and Flag-Rta can not be immunoprecipitated by each other. I also tried Flag beads, which are provided by Sigma, both of Flag-Luc and Flag-ER caught CTCF. I can not prove the interaction between CTCF and EBV Rta in TW01 inducible cell lines.

b) in co-transfected HEK293/293FT cell lines(Flag-Luc/ER/KR), CTCF can immunoprecipitate Flag-ER. However, the ratio of each plasmid is unfair, i will adjust the amount of these plasmids (may ignore Luc) and perform this experiment again.

2. to screen 12 putative target genes in CTCF/Flag-ChIP samples

利用Q-PCR的方式, 這12個genes無法在CTCF-ChIP的samples中被偵測到; 但利用一般PCR的方式, 則可偵測到JUN及H19(雖然band很微弱)

a) 在ChIP之前的sonication片段可能太短, 以至於所設計的這些將近500 bp PCR product的primer不能bind上DNA

b) ChIP的效率不好, 在抗體/磁珠/sample量的比例上該做些調整