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Questions related to Real-Time PCR
hello every body i want to design primer for my sequences but because of cross-pollination nature of my plants (Thymus sp) my sequences are heterozygot (they have SNP). I tried to design primer...
02 February 2017 9,812 4 View
In the case I do not have a standard curve.
02 February 2017 584 0 View
I am trying to figure out which C4A are long and which are short (and the same for C4B). We have a plan to do a study of complement component 4 using RT-PCR on the Biomark HD. We have decided...
02 February 2017 1,052 0 View
When performing conventional RT-PCR we did not detect TP53 expression, but when performing real time PCR with a Taqman assay for the exon 10-11 boundary, we detected expression. Maybe there´s a...
02 February 2017 7,359 1 View
Do you think the most accurate method is real-time pcr?
24 January 2017 5,428 7 View
This will be long, but I am out of ideas and require help. I am having difficulty with my real time PCR as I am obtaining multiple products in each sample. I am looking for expression of JDP2 in...
17 January 2017 9,895 6 View
I'm going to expose cells to hypoxic conditions, in order to establish the function of a synthetic promoter containing HRE's. For control sake, I want to see if cells actually developed hypoxic...
11 January 2017 6,587 3 View
Hi, I am working on circulating RNAs available in human plasma. After cDNA synthesis using TaKaRa Kit I performed qPCR (with amplicon) with GAPDH and U6 primers, but the result is a bit confusing...
02 January 2017 7,105 4 View
01 January 2017 6,628 5 View
To check the primer specificity we do run a melt curve analysis and a single curve implies the formation of a single product. But I have seen in many papers people reconfirming the specificity by...
01 January 2017 3,330 10 View
Hello I am working on rat models of acute kidney injury that induce with glycerol and ischemia reperfusion. I want to check therapeutic effects of mesenchymal stem cells of T cells subset in...
01 January 2017 5,203 1 View
Hi, I'm using the rotor gene q for my CPM assay and in the setup of the analysis I have to adjust the ramping rate to be 2˚C/min but the only way to do this is to adjust (wait for -- seconds for...
01 January 2017 8,805 0 View
01 January 2017 9,935 8 View
Hello everybody,I am optimizing transporter protein primers of coleus with actin(250bp). I used normal qPCR conditions for the optimization with annealing at 55oC and cycles 55. Most of primers...
01 January 2017 7,237 16 View
01 January 2017 6,884 4 View
Hi guys HCD test is a common test for quality control of recombinant protein in order to investigation of residual Host Cell DNA as a contamination in drugs. Considering 16srRNA repeats in E.coli...
01 January 2017 3,439 0 View
Hi everyone, I want to design a protocol to detect viral RNA of Hepatitis C virus though real time PCR with probes. I used to use a regular qPCR with primers against the 5'UTR region (most...
01 January 2017 751 0 View
Hi, I would like to get any inputs on determining the lowest viral detection limits using real time PCR. My initial trials have failed to give an amplification plot in the software, but I see...
01 January 2017 2,056 6 View
will large size 1,8kb of PCR affects ligation with 5.3kb of plasmid? I have repeated this three times; still, I'm not getting it, why it is not happening?
29 December 2016 4,758 5 View
We are going to do real time PCR with TaqMan SNP Genotyping assay to determine the genotypes of a population. However we donot have a positive control. I am wondering that is it possible to do...
22 December 2016 8,924 6 View
I am working with 16s rRNA , my amplicon size is 390 bp with 315f-806r , however I have got 350 bp , is it normal? please , if any one know answer , replay to me
12 December 2016 9,615 3 View
Hello! I'm trying to perform an optimization of qPCR for four genes of snail's nervous system (two isoforms of the same kinase and two housekeeping genes). I designed primers, checked them in...
12 December 2016 8,613 29 View
12 December 2016 9,670 6 View
Today, I set up the 1st PCR with 2.0µl templates following: 1, 5: negative control (water) 2, 6: Plasmid 1 (0.2ng/µl) 3, 7: Plasmid 2 (0.2ng/µl) 4, 8: Plasmid 3 (0.2ng/µl) Primers: gag-691F and...
12 December 2016 2,782 11 View