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 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 9,676 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 5,253 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 8,520 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 1,599 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 5,084 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 6,982 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 6,313 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 9,854 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 1,572 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 4,428 2 View
Hi RG community! I'm writing this topic because one of my friends told me he has been experiencing evaporation issues with the Applied Biosystems' 9700, Veriti and ProFlex Thermal cyclers he has...
15 March 2021 8,699 10 View
Any details regarding what you should a should not vortex would be greatly appreciated!
11 March 2021 8,382 6 View
I have recently started working on gene cloning I want to digest my PCR product and my plasmid with BamH1 and Nde1 but I am not able to digest them using 1 unit of enzyme for 10 micro gram of DNA....
11 March 2021 4,728 7 View
I am attempting to optimize my RT-qPCR to run on Taqman Fast Virus Chemistry. My efficiency when running a standard curve is only about 80%. The amplicon produced is only approx 100bp and I get...
10 March 2021 7,682 2 View
We do not have a scale to monitor the LCO2 tank weight. Is there any other way to determine when 50% of the net LCO2 has been used? If useful, I am using it to dry bacterial biofilms on glass...
10 March 2021 1,332 0 View
Dear all I hope you are having a good time. I am trying to ligate a PCR product of ~1500bp into a 6.5kbp vector (6.023kbp upon digestion). Somehow, I am not able to get colonies at all ! The...
08 March 2021 3,642 7 View
Does anyone have the experience of using Taq Man probes in the QIAGEN Rotar- Gene qPCR machine?
02 March 2021 5,699 1 View
After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...
26 February 2021 5,994 3 View
As long as I know FAM is one of the brightest used fluorophore molecule when making a qPCR test. So why Am I getting this low RFU fluorescence mixed with kind of the normal signal I would get from...
22 February 2021 6,069 1 View
Hello, Here is my experiment: I transfect in a plasmid that has been cut once (a linear piece of dsDNA) into human cultured cells. I wait 48hrs to allow the cells to "repair" the plasmid making...
11 February 2021 6,260 3 View
I am very new to quantitative PCR and need some help for this technique. For my experiment, I need to perform a quantitative reverse transcription PCR. I extract the RNA from my pellet and...
06 February 2021 10,075 4 View
The parameters of the curve all seem acceptable, typically reactions results are above 0.9852, -3.1 to -3.6 slope, and 90-110% reaction efficiency. The results seem to be what I would expect from...
05 February 2021 1,356 3 View
Hi I am having issue's with Gibson assembly, seeing as I am new in the filed of cloning, expert feedback would be very much appreciated I am trying to clone two fragments (frag 1 [185bp, 145bp w/o...
30 January 2021 1,559 2 View
I want to detection 600 kDa of high molecular weight for 4% acrylamide gel. But i could not detect that i prepared 4% acrylamide gel... Could you tell me how to prepare a 4% acrylamide gel?
26 January 2021 1,478 3 View