The dye used in BlueAmp 2X PCR Mix w/ FluoDye can be excited by blue light or UV light.
Dilute 1 part of BlueAmp DNA SafeDye with 5 parts of DNA sample and mix. Load the sample onto the agarose gel and perform gel electrophoresis.
Both Blue-Light and UV transilluminator can detect the signal.
Periodically wipe it, clean of dust, and other residue that comes from normal operation of the unit. Use a soft, lint-free cloth and deionized water. Air vents should be vacuumed to remove dust.
Before cleaning the lid heater, make sure the TurboQ is turned off, unplugged, and cooled down. Use a mild detergent to clean debris from the lid. A Kimwipe™ dipped in 70% ethanol will help remove residue from the seal. Make sure the lid is dry before plugging in the power cable.
-Absolute quantification involves the determination of the number of molecules or the copy number in the sample. It applies to the concentration of a standard sample for making a standard curve, which allows a comparison to be made with unknown samples for an estimation of their concentration.
-Relative quantification involves the comparison of differences between unknown samples and a reference gene. The test will indicate how many folds more or less it has compared to the reference gene. This is commonly used in treatment tests, to compare control genes with treated genes.
Yes, see below:
-melt curve: the melting curve traces progress as the double stranded DNA slowly degrades to single strands with increasing temperature. It can be used to determine the presence of nonspecific products after PCR cycling.
-high resolution melt curve: a high-resolution melt curve provides a quantitative analysis of the melting of double stranded DNA after PCR cycling. A high resolution melt curve can read the sample in 0.1°C increments, which means it can differentiate samples very precisely and can show differences caused by a single base pair. Only the qPCR with great thermal stability and sensitivity which has HRM-dedicated software can prepare a “high resolution melt curve”. It can be used for the analysis of SNP genotyping, gene mutation, gene methylation, microRNA… etc.
-Optical calibration: Optical calibration is only needed at first-time installation, or after the equipment has been moved to another site or position and has been reinstalled.
-Dye calibration: We recommend annual dye calibration.
However, dye calibration should be done if experimental results become abnormal.
Ct based analysis is most commonly used, but if the efficiency of your samples is low and it is difficult to reach a plateau, it may be impossible to get a correct Ct value. In such a situation, RFU based analysis which can discriminate different genes will get better results.
Yes, some enzymes are very sensitive to small changes in temperature. So, we recommend that the ramping rate be set according to the user manual recommendation for a particular enzyme.
There are four detection channels in TurboQ, and the detected fluorescence of each channel is as below.
-Ch1: FAM, SYBR Green
-Ch2: HEX, VIC, JOE
-Ch3: ROX, Texas Red
-Ch4: Cy5