I have so far disliked Taqs that come with dyes because dye concentration seems excessive to me. I have recently bought a Taq with dye-free buffer, I wonder if I can use a small bit of gel loading buffer inside my pcr reactions. Thanks!
A need to analyse specific sequences gathered from genbank in order to analyse primers matching/designing. I have used Sequencher (gene codes) in the past and I loved it. Yet prices are now...
24 April 2024 259 3 View
Having problems with dimers and an answer from Paul Rutland for another question made me think that the fact that I store my PCR mix in the fridge might be the problem. I must add that in this mix...
28 March 2024 9,886 3 View
I have a dataset of trace elements LA-ICP-MS analyses with raw data, which consists of a set of a single file for each spot analysis. Each of these files has some columns (masses) and plenty of...
24 March 2024 898 6 View
Hi all. I have some primer pairs which always produce those horrible primer dimer bright smear! Increasing annealing temperature does not solve the problem. Any suggestiopn for a PCR enhancer or...
19 March 2024 6,067 4 View
Need do look for milk bacterial pathogens using PCR. Does anyone suggest me a good protocol for DNA extraction? I have heard milk proteins are difficult to deal with aabd act as taq inhibitors
13 March 2024 402 0 View
I am making tests for diagnostic PCRs. At the moment I am trying to standardize a pcr reaction for a circa 180 bp product. I did not get positive bands so far. Interestingly, I get what appears to...
26 February 2024 2,820 3 View
We would like to preserve RNA from mammal blood samples. Unfortunately the only product I am aware is RNA Later and it is very very expensive for our limited budget. Any ideas? Thank you very much
12 December 2023 6,163 2 View
Sorry basic question. I am aware primers should be described in the 5'-3' direction in the literature. When I am ordering primers I have taken from published papers, should I use the same sequence...
18 October 2023 5,523 3 View
We are currently having problems with PCR amplifations of a ~200bp gene fragment. We cannot see any product after a 35 cycles amplification round and usually use a simple reamplification protocol:...
27 September 2023 9,212 1 View
Its manual doesn't have the conversion factor as Brookfield's does. I wish to convert rpm to shear rate but I don't know how.
08 April 2023 4,351 1 View
I got these smeared bands quite often lately. We typically run the gel at 140V with a 10-12% gel and do a wet transfer at 220 mA for 1.5 hr in cold room. We also noticed some dirty spots/dots (see...
10 August 2024 7,480 3 View
Does anyone tried to do nucleofection with AMAXA by Lonza with lower than recommended amount of buffer in the cuvettes (100 ul)?
07 August 2024 4,616 0 View
Currently, when I run SDS-PAGE, I don't see any bands at all, even though I used the same material just a day ago and it worked fine.... In our lab, we dilute the 10X running buffer to 1X and...
06 August 2024 5,373 2 View
I have been working on Red blood cell-derived extracellular vesicles as Antisense Oligonucleotide (ASO) carriers. We normally run agarose gel to quantify the loading efficiency. I used naked ASO...
06 August 2024 3,130 2 View
Hello everyone, I'm encountering an issue with my electrochemical impedance spectroscopy (EIS) measurements and would appreciate some insights. Experimental Setup: Electrodes: Gold interdigitated...
05 August 2024 3,783 2 View
I am having an issue with my gel image where my PCR product is not appearing very bright on the gel. When I perform gel extraction, the A260/280 purity value is very low. I used the Qiagen gel...
05 August 2024 9,798 3 View
I am interested to know the behavior of dyes toward light. Specifically, Blue dyes re-emit the spectrum, especially from the green zone (known as principal in LED lamps, and blue dyes are known...
05 August 2024 3,290 1 View
Is it the "elution buffer" or the "dialysis buffer"? Note: I'll be using NanoDrop OneC
01 August 2024 967 3 View
Hello everyone, I performed a PCR yesterday, and the results showed no bands on the gel. Of course, I probably missed some crucial steps, like adding my samples to the PCR strips themselves, for...
31 July 2024 2,406 6 View
We are working on biopolymeric hydrogels. Our system is highly viscous and sticky, and the gel formed are high in strength. We are unable to use pH electrode and pH strip. Please suggest an easy...
30 July 2024 942 2 View