I have a question that during primer designing I got different length of forward and reverse primer of same gene. Can I use this primer for RT-PCR?
It's no problem, but you shoud remain the same Tm value for these two primers
No problem. It's not necessary for primers forward and reverse have the same lenght.
Some useful guide for PCR primer design:
http://www.premierbiosoft.com/tech_notes/PCR_Primer_Design.html
Tm value of both the primers should be +/- 2, will work perfectly with your PCR.
There is no reason to avoid different primer lengths
Yes, you can use primers that have different lenghts
Absolutely no reason for not using a primer set having different lengths as long as their Tms dont vary much
Yes, not an issue
You can use primer set having different lengths. No problem.
06 July 2017 8,152 0 View
11 December 2016 8,539 1 View
I am trying to amplify the promoter region of 2.5 Kb of a rice gene, it is highly GC rich, but I could not amplify it even after changing primers. My primer is totally 43 bp including the 15 bp...
07 August 2016 4,841 3 View
Results of single-case research designs (i.e., n-of-1 trials) are often evaluated by visually inspecting the time-series graph and computing quantitative indices. A question our research team is...
03 March 2021 687 1 View
I have a dataset with about 80 different species. As usual, some species are very easy to identify with certainty whereas others are more difficult, which means that I am less certain of my...
03 March 2021 8,066 4 View
Hello, We would like to increase the yield of our PCR product. We are running a series of PCR reactions that is targeting ~1.1kb sequence. We begin each reaction with ~400pg of template DNA...
02 March 2021 4,029 3 View
how to calculate BEHAVIOURAL REGULATION IN EXERCISE QUESTIONNAIRE BREQ 3 ,which contain 24 questions and each question has 5 answers (0- Strongly disagree , 1-disagree, 2- Neither agree nor...
02 March 2021 811 3 View
So, I have been trying to run a pACYC PCR which will be used later on for a Gibson Assembly. However the PCR is not working. I have already tried gradient PCR and changing extension time; however...
02 March 2021 1,146 2 View
I am at the beginner level in drug design and drug discovery, and I am interested to learn more about this field. Which books are highly recommended to start from and gives me all essential...
02 March 2021 8,437 2 View
Please provide Book Title and author name
02 March 2021 9,059 3 View
We have one plant, the local people using this for controlling blood sugar, the toxicity and antidiabetic activity of the plant is not reported yet, please guide from whag should be the process ?...
02 March 2021 2,300 1 View
I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...
01 March 2021 8,544 1 View
I have to amplify a gene and my primers just reached. The Tm for Forward primer is 64.2, and that of reverse primer is 65.5. Can some one suggest how to get the best annealing temperature? Thanks...
01 March 2021 360 7 View