注:優れたPCR性能を得るには、次世代酵素Platinum II TaqホットスタートDNAポリメラーゼをお勧めします。Platinum II TaqホットスタートDNAポリメラーゼは、革新的なバッファー、高性能なTaq DNAポリメラーゼおよび優れたホットスタート技術の独自の組み合わせにより、プライマーアニーリングがあらゆる目的に対応できるように、かつPCRを迅速かつ簡単に実施できるように設計されています。
Can I purchase just the 10X PCR buffer that comes with Platinum Taq?
The 10X PCR buffer for Platinum Taq is not available as a stand-alone item. It is only supplied as part of the enzyme kit.
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.
De novo ceramide regulates the alternative splicing of caspase 9 and Bcl-x in A549 lung adenocarcinoma cells. Dependence on protein phosphatase-1.
Authors: Chalfant Charles E; Rathman Kristin; Pinkerman Ryan L; Wood Rachel E; Obeid Lina M; Ogretmen Besim; Hannun Yusuf A;
Journal:J Biol Chem
PubMed ID:11801602
'In a recent study, we showed that ceramide induces the dephosphorylation of SR proteins, a family of protein factors that regulate alternative splicing. In this study, the regulation of the alternative processing of pre-mRNA of both caspase 9 and Bcl-x(L) was examined in response to ceramide. Treatment of A549 lung ... More
Quantitative Assessment of Gene Targeting in Vitro and in Vivo by the Pancreatic Transcription Factor, Pdx1. IMPORTANCE OF CHROMATIN STRUCTURE IN DIRECTING PROMOTER BINDING.
Authors: Chakrabarti Swarup K; James Joshua C; Mirmira Raghavendra G;
Journal:J Biol Chem
PubMed ID:11825903
'The transcription factor Pdx1 is expressed in the pancreatic beta-cell, where it is believed to regulate several beta-cell-specific genes. Whereas binding by Pdx1 to elements of beta-cell genes has been demonstrated in vitro, almost none of these genes has been demonstrated to be a direct binding target for Pdx1 within ... More
Angiotensin II inhibits human trophoblast invasion through AT1 receptor activation.
Authors: Xia Yang; Wen Hong Y; Kellems Rodney E;
Journal:J Biol Chem
PubMed ID:11983698
Trophoblast implantation depends, in part, on the controlled production of plasmin from plasminogen, a process regulated by plasminogen activators and plasminogen activator inhibitors. We have determined that angiotensin II (Ang II) stimulates plasminogen activator inhibitor-1 (PAI-1) synthesis and secretion in human trophoblasts in a time- and concentration-dependent manner. Our results ... More
Sensory nerves determine the pattern of arterial differentiation and blood vessel branching in the skin.
Authors: Mukouyama Yoh suke; Shin Donghun; Britsch Stefan; Taniguchi Masahiko; Anderson David J;
Journal:Cell
PubMed ID:12086669
Nerves and blood vessels are branched structures, but whether their branching patterns are established independently or coordinately is not clear. Here we show that arteries, but not veins, are specifically aligned with peripheral nerves in embryonic mouse limb skin. Mutations that eliminate peripheral sensory nerves or Schwann cells prevent proper ... More
Translational regulation of BACE-1 expression in neuronal and non-neuronal cells.
Authors:De Pietri Tonelli D, Mihailovich M, Di Cesare A, Codazzi F, Grohovaz F, Zacchetti D,
Journal:Nucleic Acids Res
PubMed ID:15034149
As the main beta-secretase of the central nervous system, BACE-1 is a key protein in the pathogenesis of Alzheimer's disease. Excessive expression of the protein might cause an overproduction of the neurotoxic beta-amyloid peptide. Therefore, a tight regulation of BACE-1 expression is expected in vivo. In addition to a possible ... More