I need to add constraint on moment to fit data in FullProf. I set NMAGC to 1 and added constraints as followings.
! Nsym Cen N_Clat N_Ant NMAGC
4 0 0 0 1
!
! Soft moment constraints:
CATOM, Moment, Sigma
Eu1 5 1
Eu2 5 1
Eu3 5 1
!
But I received error message "Error in Shubnikov operator: (A2, 2 reals): CATOM, Moment, Sigma."
Any advice where I made silly mistake? Thanks so much in advance.
Othman Mueen Mohammed
Denaturation Step:The denaturation step typically involves heating the sample to a high temperature (usually around 94-98°C) to separate the double-stranded DNA into single strands. The duration of this step is usually 30 seconds to 1 minute. Ensure that the denaturation temperature and time are sufficient to completely separate the DNA strands without causing damage to the DNA polymerase. Annealing Step:The annealing step involves cooling the sample to a temperature that allows the primers to bind to their complementary sequences on the single-stranded DNA. The annealing temperature is typically 5-10°C below the melting temperature (Tm) of the primers. The duration of the annealing step is usually 30 seconds to 1 minute. Optimize the annealing temperature and time to ensure specific primer binding and minimize non-specific amplification. Extension (Elongation) Step:The extension step involves heating the sample to a temperature optimal for the DNA polymerase to synthesize new DNA strands. The extension temperature is usually around 72°C. The duration of the extension step depends on the length of the DNA fragment being amplified. A general rule of thumb is to allow 1 minute per 1 kb of DNA. Cycle Number:The number of PCR cycles should be optimized to ensure sufficient amplification of the target DNA without reaching a plateau phase. Typically, 25-35 cycles are used, but this can vary depending on the initial amount of template DNA and the efficiency of the PCR reaction. Final Extension:After the last cycle, a final extension step at 72°C for 5-10 minutes is often included to ensure that any remaining single-stranded DNA is fully extended. Optimization and Validation:Perform optimization experiments to determine the optimal conditions for each step. This may involve varying the temperatures, times, and reagent concentrations to achieve the best results. Validate the optimized conditions by running control experiments and checking the specificity and efficiency of the PCR reaction.
o add the correct moment constraint in PCR, you need to ensure that the timing and conditions of each step in the PCR cycle are optimized for your specific experiment. Here are some key points to consider:
0 votes
0 thanks
- Badges
- Science topic
- Similar topics
- Social Psychology
- Attitude
More Qiang Zhang's questions See All
Similar questions and discussions