In vitro method for producing large amounts of specific DNA or RNA fragments of defined length and sequence from small amounts of short... | Contact experts in PCR to get answers
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Questions related to PCR
can someone explain me how to determine LOD and LOQ value of qRT-PCR using probit regression ? i've calculated LOD and LOQ value of my qRT-PCR result using regression linear, but in some...
16 August 2021 9,391 2 View
For the optimization of primers for bisulphite sequencing, I extracted DNA from blood which was followed by its bisulphite conversion. Through gradient PCR, all primers were standardized...
15 August 2021 4,766 6 View
Which one is the best option available for sealing PCR plates? 1. Sealing films or 2. Sealing aluminium foils or 3. TPE or Rubber Mats or 4. Lids I am having TPE Mats. So, can I use the same again...
09 August 2021 6,669 3 View
I am trying to do a dft calculation in gromacs but don't know the code. Can anyone provide me the detailed code of the same.
06 August 2021 7,032 0 View
Our target aptamer sequence is small (78bp). We are facing a problem during cloning. Is cloning necessary for sequencing of our target aptamer (DNA) or any other way is present? Any suggestions...
05 August 2021 1,012 3 View
We are trying to purchase a suitable set of primers and restriction enzymes for the qualitative diagnosis of of chronic myeloid leukaemia using PCR and restriction fragment length poly,orphism....
05 August 2021 6,827 3 View
Hello everyone .I’m trying to do TETRA ARMS PCR. My outer Forward and Reverse primers have TM: 58 and TM: 50 and inner Forward and Reverse primers have TM: 52 and TM: 48. How can I setup at...
04 August 2021 1,836 1 View
What is the meaning of having results when using a specific primer for E.coli gene, for example, in klebsiella pneumonia.. The gene product is present in both, but is it normal that the gene is...
03 August 2021 7,776 5 View
I am doing site directed mutagenesis using stratagene-QuikChange II Site-Directed Mutagenesis Kit. The protocol and primer design I'm following are given in the link below. I am not getting any...
02 August 2021 845 9 View
Hello, I’m studying population genetic structure of fish in Thailand. by DNA sequencing using mtDNA COI. How I can calculate the appropriate number of samples from 6 populations. Thank you in...
26 July 2021 8,359 3 View
I have been trying to perform a single amino acid mutation using a the GENEART site-directed mutagenesis system. I isolated the plasmid, fixed its concentration at 50ng/ml for the mutagenesis PCR...
25 July 2021 3,775 4 View
We use a manual droplet generation microfluidic device to generate droplet. We use syber green dye for this purpose (PCR). Is this dye appropriate for ddPCR? What ratio of DNA, primer and...
19 July 2021 4,487 4 View
I have made detection for microsatellite SSR markers in patient with plasmodium... for statistical analysis, I have used genalex 6.5 ... I still need to make confirmation with another...
15 July 2021 7,851 6 View
Hi everyone, I have a very troublesome time when I trying to build the construct of my gene. Since the gene sequence of my target insert is toxic due to the leaky expression, I used the competent...
14 July 2021 7,498 15 View
Our in-house mycoplasma testing flagged both my media types as positive for mycoplasma, but the cell line media samples I provided all tested negative. The tests were performed on different days...
12 July 2021 8,317 4 View
I am trying to amplify 16S regions from insect gDNA samples. I am getting the target bands with low intensity. I am following standard NEB protocol for 25ul reaction tube. 7ul of PCR product was...
08 July 2021 8,641 24 View
Hi, As part of my research work, I require to perform semipreparative chromatography to isolate phenolic rich fractions from crude seaweed extracts (mostly brown algae). The majority of these...
08 July 2021 4,783 0 View
I have performed a qPCR test to check my primer efficiency. I got the rough data now which I have opened with Design and Analysis software 2.5.1. It is quite confusing and I am relatively new in...
01 July 2021 6,450 2 View
While working on culturing of Aspergillus Flavus, I got this dark gray fungus on one of the plates. I wish to know the name and the colony characteristics or identification factors for this fungus.
27 June 2021 2,170 3 View
I am doing an allele-specific PCR to detect a point mutation in the DNA of Aedes aegypti, when I used a tm temperature of 55 °C, I observed some PCR products in negative control, and some...
26 June 2021 9,547 5 View
I'm trying to isolate DNA from fibroblast cultured cells without using a kit (with lysis buffer and prot K), but after PCR amplification, no products is shown on the agarose gel. However, the same...
15 June 2021 5,979 2 View
I am trying to design primers for some miRNAs but I keep getting errors pertaining to my minimum and maximum PCR product size. Any help?
14 June 2021 4,925 12 View
I extracted DNA from some kombucha products with a manual protocol, with one final step using phenol, for 16S and ITS sequencing. However, there is a problem for the majority of my samples in PCR...
13 June 2021 9,614 3 View
Do we need to do a J-model test before preparing the phylogenetic tree from RAxML or after it is prepared? In both the conditions what are the values of different parameters we need to add. Please...
12 June 2021 4,247 2 View