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
Hi, Please anyone already prepared the mix for Standard PCR? But without using a commercial kit. It is for staphylococcal enterotoxins genes. I am looking for the concentrations of each...
02 February 2020 569 8 View
This is the image of DNA agarose gel where I have loaded the PCR reaction mixture. I see a trail in my DNA gel that starts from the lane. It can't be the non-specific PCR products as the trail is...
02 February 2020 2,819 2 View
Hello all! I am currently looking at either using Exo-CIP rapid clean up or ExoSAP-IT (non-express). I was hoping to get some people's opinions on these two products. It looks like Exo-CIP express...
02 February 2020 3,813 3 View
I am using the following protocol from a CSH lab manual edited by Doudna on CRISPR, to attempt to retarget sgrna scaffold in a plasmid. but....I get DNA stuck in the well, not migrating....
02 February 2020 6,424 3 View
Currently trying to amplify my 15kb plasmid with specific primers I designed for the desired ends of the linearized fragment. I'm currently using Phusion polymerase by NEB, and using the suggested...
02 February 2020 9,463 3 View
I am trying to find a good qRT-PCR primer set for a gene that has low expression in our cell lines. As you see in the attachment I have a very sharp one peak when I used cDNA from overexpression...
02 February 2020 7,774 4 View
Dear all, as luck would have it, I reached for my PureLink PCR Micro kit and the wash buffer totally evaporated. Does anyone know the composition of the wash buffer in the PureLink PCR Micro kit?...
02 February 2020 290 1 View
What affects the viscosity of PCR reactions? I mean the PCR solution, not the reaction chamber or tube. For example, may be temperature, concentration of ion or DNA. I want to make quantitative...
02 February 2020 9,328 3 View
I am doing qPCR experiments, I am making the reaction with same water for samples, NTC, however negative control is also giving ct
02 February 2020 7,194 3 View
I tried to identify 4039bp mRNA sequence by dividing the whole sequence into 6 fragments and designing primers for each of them. But two fragments in this sequence couldn't amplified by PCR...
02 February 2020 10,066 3 View
Hello, When I run my PCR products along with no template control, I get a band at the desired product size even for my no template control. This is happening for PCR products of varying sizes...
02 February 2020 3,149 6 View
I am using EA.hy926 endothelial-derived cells. I have tried multiple housekeeping genes but they are inconsistent even within the controls. So is there any particular housekeeping gene to use with...
02 February 2020 3,289 2 View
As an example, I suppose to keep the cycle 1 in 22 C for 5 mins while I kept it for 5 seconds.....42 C for 30 minutes while i only kept it for 3 seconds. I guess when I was entering the numbers...
02 February 2020 1,139 3 View
I'm looking for SNP variation in a population and in an article I found that the primer they use is really close (3nt) away to the SNP target, so I was wondering if there's a possibility that...
02 February 2020 4,933 3 View
Hi, I want to generate a HA-tagged fusion protein (HA-tag at protein C-terminus) during transient expression in plants. I'm choosing a PCR method over restriction digestion. Strategy is, a regular...
02 February 2020 8,035 1 View
i would be much grateful to have an opinions based on Nepalese condition. is this topic appropriate to start a UPA In bachelor level.
02 February 2020 8,750 2 View
I am using Psy I (is-schizomer of Asp I) and my PCR product didn't get in digested form. Instead I see the only band on the gel is that of my PCR product (i.e 900b.p). I use enzymes from...
02 February 2020 2,120 2 View
I am having trouble amplifying a sequence from cell-free DNA. I tested this primer before with DNA from a cell line, and it worked fine. However, when I tried PCR on human cell-free DNA, nothing...
02 February 2020 6,837 4 View
I am genotyping by amplifying a segment from gDNA. I had non-specific bands in my PCR so I did a gradient annealing temp for 30 cycles. Weirdly, the lowest temperature actually seems to give fewer...
02 February 2020 6,266 4 View
how to increase the yield of PCR product ?
02 February 2020 7,337 2 View
I don't really have access to make a complicated process for this, but I was wondering if there was a way to conduct PCR on an entire group of cells if I already have primers specific to the...
02 February 2020 1,657 3 View
I am trying to design lamp PCR primers and I am really confused about their annealing temperatures. Annealing temp for the same region in PCR is 58 degrees C but in lamp PCR primer designing...
02 February 2020 7,177 3 View
Hello, I am trying to look at the efficiency at which a Gene is cut after tamoximen treatment. Thus we got a set of primers that can tell us weather the Gene is cut or not. We wanted to measure...
02 February 2020 7,030 3 View
I have been beyond meticulous with NEB Builder, and it has never once worked. I have had success with OE PCR, but NEB Builder fails totally, every time. I get nothing but smear and concatenation...
02 February 2020 9,100 9 View