How to Choose the Right PCR ReagentsTanuja Koppal, Ph.D
After adding the nucleic acid to be detected, the reaction system performs a fluorescent PCR amplification reaction. The time is slightly different depending on the PCR reagents and instruments used. Generally,it takes 90-120 mi...
The program on the thermal cycler will be similar to that of the PCR. Cycle sequencing clean-up A sequencing clean-up step is then performed prior to capillary electrophoresis. This is different from the PCR reaction clean-up; this step is need...
Only primers and sample of DNA source need to be added to the PCR master mix to run reactions. Bonus tip: Depending on the genes you are detecting, you may be able to include multiple primer sets in a single reaction (i.e., multiplex PCR). In the examp...
Each restriction enzyme has optimal reaction (assay) conditions and different conditions for long term storage. The recommended assay and storage conditions are both determined by the manufacturer to provide the user with the highest activity, best fidelity and greatest stability for each enzyme. Factor...
Polymerase Chain Reaction is the in-vitro method of making multiple copies of a DNA sequence. The three main phases of PCR are Denaturation, Annealing, and Elongation.Answer and Explanation: During PCR, the function of DNA polymerase is the add the dNTPs (deoxyribonucleotide triphosphates) to ...
When real-time polymerase chain reaction came out during the 90s the words kinetic PCR analysis or real-time PCR were most commonly used [1][2][3]. Also, real-time quantitative RT-PCR was used to indicate RT-qPCR [4]. When real-time PCR became more common in the 2000s it was more...
Obviously, a perfect primer set will have a primer efficiency of 100%. In other words, for every PCR cycle, the number of copies of the PCR product will double in size during the logarithmic phase of the PCR reaction. To get a 100% primer efficiency for all of your primer sets is hig...
The instrument specific properties conceived to affect the PCR efficiency are the reaction container, which here in most cases were plastic microtiter well plates, that influences the rate and efficiency of heat transfer, the heating and cooling capacities, which may induce thermal over- and ...
Once set up, a function such as new_tip was used to get new tips in sequential order starting from the right bottom of the pipet box (Fig. 5A). Whereas the eject function ejected the pipet tip by applying force directly to the pipet cone head from the tip ejector trigger (Fig. 5H)...