SYBR® Green Realtime PCR Master Mix - Toyobo

www.toyobo.co.jp/e/bio[ 7 ] TroubleshootingSymptom Cause SolutionIncorrect mode setting of detector Confirm the detector setting.for the fluorescence dye.Incorrect setting for data collection. Confirm the data collection setting.No amplificationVariation in detectionSignals in blankreactionsIncorrect setting for sample position.Inappropriate concentration ofprimers.Inappropriate design of primers.Inappropriate cycle conditions.Low purity or quality of samplesFailure or malfunction of the deviceLow quality sample DNA.Inappropriate concentration ofprimers.Inappropriate design of primers.Inappropriate cycle conditions.Variation of dispensed volume.Contamination ofamplicons or sample DNAs.Detection of non-specificamplification.Reposition the sample tubes.Optimize the concentration of primers accordingto instructions (see [6]).Confirm specificity and Tm value of the primers.Confirm the primer Tm value. A lower annealingtemperature increases amplification efficiency.For GC rich targets, an elongated denaturationtime might be effective.Check the purity or quality of sample DNA.Check the device.Repurify the sample DNA by phenol/chloroformextraction and ethanol precipitation, or othermethod.Optimize the concentration of primers accordingto instructions (see [6]).Confirm specificity and Tm values of the primers.Confirm the Tm values of the primers.Increase the reaction volume.Use fresh PCR grade water, and remake theprimer solution and master mix.Optimize the primer and cycle conditions.JAPANTOYOBO CO., LTD.Tel(81)-6-6348-3888www.toyobo.co.jp/e/biotech_osaka@toyobo.jpCHINATOYOBO Bio-Technology, CO., LTD.Tel(86)-21-58794900.41407FOR RESEARCH USE ONLY. NOT FOR HUMAN OR DIAGNOSTIC USE.