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
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 832 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 8,348 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 3,762 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 4,474 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 7,839 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 7,484 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 8,305 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 8,630 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 4,762 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 6,440 2 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 9,535 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 5,969 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 4,912 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 9,602 3 View
I m a first year student so this question might seem a little dense. but currently I ran PCR analysis of some gene. I got the potential interesting genes from the microarray data from the geo...
11 June 2021 9,663 3 View
I would like to use an exogenous internal positive control in my qPCR reactions (probe-based with gDNA). The IPC protocol states a 60C anneal/extend step, but my target assays require this step to...
07 June 2021 5,234 5 View
I am facing difficulties in cloning a 2.5kb DNA fragment into a vector (pGreen0029). My plates were designed as +ive = pGreen0029, -ive = competent cells (cc), L1-3 = ligation reactions performed...
25 May 2021 8,503 3 View
We are trying to transfect plasmid DNA into the THP-1 and primary monocytes. However, transfection efficiencies of lipofectamine 2000 and Genjet were not enough to perform subsequent downstream...
22 May 2021 1,584 1 View
Hi Everyone; Fİrst of all, i know the basics of a primer-probe design. What i'm looking for is detailed workshops or courses that i can learn the tricks for a perfect PCR reaction. If anyone here...
20 May 2021 5,070 6 View
Could you please recommend to me some manufacturer of Lyo-ready RT qPCR Master Mix, ideally accessible from Europe? I am aware of ThermoFisher and MeridianBioscience. Thank you very much!
20 May 2021 6,971 2 View
Discuss the impacts of local and global change challenges on Forest Landscape Restoration 1.Climate Change 2.Food Security 3.Biotechnology and Synthetic Biology
11 May 2021 6,303 8 View
The instrument fires up and runs good, however when you shoot the laser no MS shows in the buffers and it just freezes afterwards. It seems like the instrument is either not sending the data due...
29 April 2021 9,842 0 View
My lab is working with primary cells from the patient and we would like to check if our cells are infected with HIV, HBV, HCV, etc by PCR. We would like to know how many cells do we need for this....
19 April 2021 1,555 2 View
Hi everyone, I am interested in performing fingerpitng DNA tests with ISSR and RAPD markers. Is a special purification of the primers needed (PAGE or HPLC) ? Can the type of purification affect...
17 March 2021 4,421 2 View