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Questions related to PCR Analysis
Hi everyone, I would like to know that if my normalized gene " 18srRNA" was detected at ct=9 but my interested gene was detected at ct=29, I would like to know that why is the low ct not good for...
23 October 2015 7,751 3 View
I am going to used methylated sensitive restriction enzymes combine with PCR to analysis methylation status of cytisines in a plant gene. While restriction site of HpaII/HspI can be determined...
10 October 2015 6,220 0 View
I've been having trouble with a PCR reaction. I ran two PCRs with the same DNA samples, the only different being the primers. The first PCR (labeled Zip14 WT) worked fine. The second PCR (labeled...
10 October 2015 6,419 11 View
can I amplify FOXO1 and RUNX2 with proof reading polymerase such as high-quality Taq polymerase. Then, incubate the PCR product with ordinary Taq polymerase to generate A overhang Also, I would...
10 October 2015 5,186 3 View
What information can be determined by using cDNA as opposed to genomic DNA as the template for a PCR reaction?
10 October 2015 2,332 0 View
The length of the construct is 5.6 KB including gene. PCR reaction set up... Q5 master mix- 12.5 ul 10uM forward primer-1.25 uL 10um reverse primer-1.25 Template-50 ng PCR CYCLE- initial...
10 October 2015 5,527 3 View
Asymmetric PCR of oligonucleotide of less than 100 bases reverse primer and forward primers of ~27 bases each How to confirm that the product is the desired ssDNA Any kits for purification and...
10 October 2015 2,280 9 View
I recently started working with LAMP (Loop-mediated Isothermal Amplification) using target DNA. I designed 4 primers using PrimerExplorer V3. I use LAMP Kit by Lucigen, Inc. For some reason I do...
09 September 2015 6,100 3 View
I ask because most of them changes its expression under physical exercise. I checked it for HPRT 2 cycles of change , ACTB much more, GAPDH much more. Of how to treat articles when e.g. these...
09 September 2015 6,040 2 View
I have two viruses that are fairly similar. The design of the primer relied basically in minimum differences since these viruses are too similar. I could make the assay work under SYGR assay but...
09 September 2015 1,495 3 View
Most of the literature available determining the expression of mTORC1 reports only western blot analysis. Could not find any PCR analysis
09 September 2015 5,117 5 View
I've faced a problem during my work.In electrophoresis of ureC gene PCR product electrophoresis H pylori the the weight of gene band is 100bp more than the positive control weight while I observe...
09 September 2015 6,692 2 View
I had weak PCR bands for Adeno virus and when I did a nested PCR using the same primer and the pcr products I got no bands, what is my mistake? Adeno virus nested-pcr Nested-PCR
09 September 2015 7,421 4 View
Does anyone know how to combine run data from multiple plates into an analysis of a single experiment in the software provided by BioRad for real-time PCR analysis (CFX Manager -- we are using...
21 August 2015 9,220 6 View
- sample 9 has a negative result using exactly the same mastermix I used for the negative control. - used a 100 bp molecular weight ladder - negative control: water - 9-14 are the samples
08 August 2015 4,753 10 View
08 August 2015 9,209 7 View
Hello, in my PhD thesis I have to use ChIPqPCR to study the differences in plant's chromatin (control vs infected). To do so, I use some antibodys that bound to widely recognised histone...
08 August 2015 2,549 25 View
Actually I need to make a construct for protein localization and I don't have any idea how to stitch so many fragments together. Can anyone suggest me how to do that starting from primer designing...
08 August 2015 979 4 View
Hi, I want to amplify a large gene
08 August 2015 4,016 4 View
Say you have three fragments that have overlaps with each other. During PCR, I'm assuming that since these fragments could physically overlap with each other they will anneal. My question is will...
08 August 2015 2,242 4 View
We are having problems with the generations of mouse monoclonal antibodies (MAbs). We have routinely produced MAbs before, but lately started having problems. Basically, we have hybridomas in more...
08 August 2015 9,015 9 View
Hi everyone, I have been working on real-time quite alot. Recently, I performed an experiment with 3 repeats. For me, repeat means doing same experiment again, but at different time. So, I did...
08 August 2015 3,108 5 View
My PCR product appear very faint with primer dimer in each samples , I tried to increase annealing temperature , the extension time and the initial denaturation temperature as recommended in PCR...
08 August 2015 5,964 16 View
I am currently trying to detect bacterial 16S rRNA DNA in human tissue to ultimately sequence the results. I conducted PCR on diluted samples (original concentration of DNA was between...
08 August 2015 9,259 6 View