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。
How do you recommend storing proteinase K?
We recommend storing it at 4°C.
How do I isolate DNA from single bacterial colonies before PCR?
Scrape a single colony from the surface of an agar plate and transfer to 12 µL of SCL solution [10 mM Tris-HCl (pH 7.5), 1 mM EDTA, 50 µg/mL proteinase K] and incubate for 15 min at 55 degrees C. Inactivate proteinase K for 15 min at 80 degrees C and add 20 µL of deionized water. Centrifuge at 12,000 x g for 3 min. Transfer the supernatant to a new tube.
引用および参考文献 (2)
引用および参考文献
Abstract
Improved cloning efficiency of polymerase chain reaction (PCR) products after proteinase K digestion.
Authors: Crowe J S; Cooper H J; Smith M A; Sims M J; Parker D; Gewert D;
Journal:Nucleic Acids Res
PubMed ID:2011503
Not available
Regulation of calreticulin expression during induction of differentiation in human myeloid cells. Evidence for remodeling of the endoplasmic reticulum.
Authors: Clark Robert A; Li Sen-Lin; Pearson Doran W; Leidal Kevin G; Clark Joshua R; Denning Gerene M; Reddick Robert; Krause Karl-Heinz; Valente Anthony J;
Journal:J Biol Chem
PubMed ID:12065601
Induction of differentiation of HL-60 human myeloid cells profoundly affected expression of calreticulin, a Ca(2+)-binding endoplasmic reticulum chaperone. Induction with Me(2)SO or retinoic acid reduced levels of calreticulin protein by approximately 60% within 4 days. Pulse-chase studies indicated that labeled calreticulin decayed at similar rates in differentiated and undifferentiated cells ... More