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
1,986 views 3,271 posts
Questions related to PCR
I use end-point PCR cleanup product as a template for my Real time PCR standard curve studies (10-fold dilutions). My slope always ranges between -3.7 to -3.9, instead of an ideal 3.3. What could...
06 March 2020 6,199 3 View
I have a query. I have set a 60ul volume of reaction volume of PCR divided into 10 ul small volumes. After PCR is done , I find a reduction in volume from 10 ul to around 8 or 6 ul in some of...
03 March 2020 8,405 4 View
I'm trying to purify my pcr product by using QIAEX II Gel extraction kit. According to the protocol, firstly i need to add buffer QX1 to the tube (contain gel slice that already been cut)...
28 February 2020 7,889 9 View
Hello, It was easy to get the full length of SbBDAH1 (1520 bp) & SbBADH2 (1517 bp) from cDNA using dream taq from thermo scientific, although there were non specific targets and primer...
28 February 2020 5,951 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...
22 February 2020 4,512 3 View
I am currently working in an isothermal amplification technique called PSR ( Polymerase spiral Reaction). I have followed the reaction mixture from previous articles but i am getting primer dimers...
16 February 2020 2,914 3 View
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...
09 February 2020 4,580 6 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...
03 February 2020 7,687 3 View
Hello, I would like to replace some amino acids in a protein but keep it functional. How can I assess the effect computationally? Could you advise some algorithm, preferably with a web...
03 February 2020 3,189 2 View
I have recorded X-ray diffraction (XRD) patterns of the Ca-ZrO2 (Powder and Ceramics) using Cu-Kα radiation (λ = 0.154 nm). The sample contains only the tetragonal phase. I want to refine using...
02 February 2020 6,712 3 View
I am doing research on the chemotoxic assay in C. elegans. One of the strains I used has a significant difference between RNAi and knockout strains. The RNAi strain is significantly different...
02 February 2020 8,733 3 View
I am doing genotyping to identify mutants. i use HD2B gene primer which are not working very efficiently. they work some times for some of the samples to barely amplify the gene (see figure...
02 February 2020 7,887 7 View
02 February 2020 8,210 3 View
Does Autoclaving and gamma irradiation remove DNA enough so that no DNA fragment will be amplified in PCR?
02 February 2020 3,481 3 View
Hi everyone, i am doing mycoplasma detection by doing Pcr and im doing PCR 10ul reaction, my primer concentration os 2pmol and vol is 0.1ul. As i dont have pipette for 0.1ul i am thinking of...
02 February 2020 8,153 3 View
Hello PCR experts, I am attempting to optimize H3 PCR for my crustaceans, but keep running upon double bands in my PCR product. The attached pictures shows a temperature gradient from 48C to 60C...
02 February 2020 2,311 4 View
We amplify genes by PCR using plasmid DNAs as templates. The downstream is to synthesize RNAs by in vitro transcription (IVT) using PCR products as IVT templates. The PCR products need to be...
02 February 2020 3,143 6 View
Hi everyone, I want to amplify soil fungal ITS2 region. But I tried three times with concentrated DNA, diluted DNA, the gel picture always showed double bands. Although I increased the anneal...
02 February 2020 6,380 3 View
HI there, I need some assistance in figuring out why my DNA will not run down my agarose gel. Here is my experimental pipeline 1: Ligate a PCR product with custom adapters 2: Run Ligation...
02 February 2020 6,005 25 View
A primer is a short single-stranded nucleic acid utilized by all living organisms in the initiation of DNA synthesis. The enzymes responsible for DNA replication, DNA polymerases, are only capable...
02 February 2020 9,230 8 View
Hello, I am looking for an alternative for Seqeuncher since our license is running out and wanted to see what people's opinion are for Sanger sequence editing. I know there are a lot out there,...
02 February 2020 9,883 1 View
I have 10 rice samples which I'm testing with housekeeping primers. 9 of them are supposed to be transgenic. And 1 is wildtype. I did a PCR with all of them, in which I had a band with all the...
02 February 2020 9,538 3 View
Hello, I am a user of QuikChange II XL Site-Directed Mutagenesis Kit (Agilent) and I have performed SDM succesfully several times. I always introduce single nucleotide changes using a pair of...
02 February 2020 9,383 4 View
I want to get a PCR product that has an NH2 group at the 5'end, so I designed primer with linked NH2 group at 5'end. However, it is difficult to get a PCR product. Is there a paper or any idea how...
02 February 2020 4,195 3 View