Hi, I want to make my own sybergreen kit using syber green dye because of cost.
I search abouth it and see the subjects dates are very old and a lot of researcher talked abouth this, they said "I tried but its not stabile."
Do anyone know abouth good protocol to make our qpcr cybergreen(or otherflorecent)
Final question, can we use SYBR™ Green I Nucleic Acid Gel Stain, 10,000X for qpcr
thank you
There's no protocol. Just dump it in there at 1x final reaction concentration. Keep in mind that your mastermix has an x factor too. So if you're making 1,000 ul of 5x mastermix, you'd add 0.5 ul of 10,000x Sybr green, not 0.1 ul. Make sense?
As for stability, we're using a Sybr Green derivative called EvaGreen. Like most dyes, it's sensitive to light, and probably warm temperature. We kept our EvaGreen-mastermix at -20 for at least a year now, and it's just fine.
John Schloendorn I'm sory because I couldnt be clear I understood from your answer Dr.
actualy I wanted to know how can I make my own qpcr master mix
Sure, you can make your own diluted stock. I also used EvaGreen instead of SYBRGreen (better signal, less noise).
I'd aliquot out single-use sized portions and had great success.
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