Applied Biosystems 7900HTの384-well blockで、FastモードのTaqMan Real-Time PCRを行う場合、User Bulletin(P/N 4369584) " Performing Fast Gene Quantitation with 384-Well Plates"に操作の詳細な指示がまとめられていますので、参照してください。
My oligonucleotide does not appear to be the right length when I checked by gel electrophoresis. Why is this?
Oligos should be run on a polyacrylamide gel containing 7 M urea and loaded with a 50% formamide solution to avoid compressions and secondary structures. Oligos of the same length and different compositions can electrophorese differently. dC's migrate fastest, followed by dA's, dT's, and then dG's. Oligos containing N's tend to run as a blurry band and generally have a problem with secondary structure.
The primers I am using worked for PCR initially, but over time, have stopped working. What happened?
Primers should be aliquoted for single use before PCR set-up. Heat just the aliquoted primers to 94 degrees for 1 min. Quick chill the primer on ice before adding to the PCR reaction. Some primers may anneal to themselves or curl up on themselves.
I don't see a pellet in my oligo tube order. Should I ask for a replacement?
The drying method dries the primer in a thin layer along the sidewalls of the tube instead of the bottom, therefore a pellet is not always visible and should still be ready to use.
There is a ball-shaped pellet at the bottom of my oligo tube. What is this and can I still use my oligo?
If the oligo was overheated, it will appear as a ball-shaped pellet attached to the bottom of the tube. This should not affect the quality of the oligo, and the oligo should be readily soluble in water.
Sp3 Represses Gene Expression via the Titration of Promoter-specific Transcription Factors.
Authors: Kennett Sarah B; Moorefield K Scott; Horowitz Jonathan M;
Journal:J Biol Chem
PubMed ID:11773047
'We have determined previously that Sp3 encodes three distinct gene products as follows: a full-length protein (Sp3) that is an activator of transcription and two isoforms (M1 and M2) derived via internal translational initiation that function as transcriptional repressors. To identify amino acids and functions required for transcriptional repression, we ... More
Both DNA and histone fold sequences contribute to archaeal nucleosome stability.
Authors: Bailey Kathryn A; Marc Frederic; Sandman Kathleen; Reeve John N;
Journal:J Biol Chem
PubMed ID:11751933
'The roles and interdependence of DNA sequence and archaeal histone fold structure in determining archaeal nucleosome stability and positioning have been determined and quantitated. The presence of four tandem copies of TTTAAAGCCG in the polylinker region of pLITMUS28 resulted in a DNA molecule with increased affinity (DeltaDeltaG of approximately 700 ... More
Nuclear factor-kappa B directs carcinoembryonic antigen-related cellular adhesion molecule 1 receptor expression in Neisseria gonorrhoeae-infected epithelial cells.
Authors: Muenzner Petra; Billker Oliver; Meyer Thomas F; Naumann Michael;
Journal:J Biol Chem
PubMed ID:11751883
'The human-specific pathogen Neisseria gonorrhoeae expresses opacity-associated (Opa) protein adhesins that bind to various members of the carcinoembryonic antigen-related cellular adhesion molecule (CEACAM) family. In this study, we have analyzed the mechanism underlying N. gonorrhoeae-induced CEACAM up-regulation in epithelial cells. Epithelial cells represent the first barrier for the microbial pathogen. ... More
Methylation-mediated silencing of TMS1/ASC is accompanied by histone hypoacetylation and CpG island-localized changes in chromatin architecture.
Authors: Stimson Krista M; Vertino Paula M;
Journal:J Biol Chem
PubMed ID:11733524
'Aberrant methylation of CpG-dense islands in the promoter regions of genes is an acquired epigenetic alteration associated with the silencing of tumor suppressor genes in human cancers. In a screen for endogenous targets of methylation-mediated gene silencing, we identified a novel CpG island-associated gene, TMS1, which is aberrantly methylated and ... More
Functional coadaptation between cytochrome c and cytochrome c oxidase within allopatric populations of a marine copepod.
Authors: Rawson Paul D; Burton Ronald S;
Journal:Proc Natl Acad Sci U S A
PubMed ID:12271133
'Geographically isolated populations may accumulate alleles that function well on their own genetic backgrounds but poorly on the genetic backgrounds of other populations. Consequently, interpopulation hybridization may produce offspring of low fitness as a result of incompatibilities arising in allopatry. Genes participating in these epistatic incompatibility systems remain largely unknown. ... More