Can I insert sequence (200bp-800bp) in pUC57 plasmid for PCR and qPCR applications??

More Luis A. Salcedo Mejía's questions See All

The incubation with isopropanol to -20°C overnight increase the ARN recuperation?

Hi all, I did read few papers that write protocols with overnight incubation with isopropanol for increase RNA recuperation, but the handbook (f.e. qiazol) does not mention it, only an incubation...

01 February 2017 1,322 5 View

What statistical technique I use and how I use it to analyze a comparison between a native strain and control strain for the solubilization ofcopper?

evaluated the bioleaching of a native strain compared with a control strain, have variables (cell concentration,% solubilization of copper, time) and the taking of the data is that starting from a...

03 April 2016 8,728 1 View

What is the exactly definition of metagenomics??

I work in Biomining and have a question.Performed from a liquid culture using molecular identification 16S rRNA is metagenomics? thank you very much!!

01 February 2015 7,502 1 View

Is it effective to determine iron II with potassium dichromate in the presence Iron III?

I realize experiments of bioleaching of polymetallic ores with bioleaching consortia, and want to determine iron (II). My protocol is: To 1mL sample (aprox pH 2- 5) add 1mL solution H3PO4:H2SO4...

10 November 2014 5,267 13 View

When using heap leach or stirred tank bioreactors, does it depend on the mineral to use?

There are many processes / patents using bioleaching of sulphide ores or oxidized copper / gold, using both batteries as bioreactors, but this will depend on that? The percentage of precious metal...

05 June 2014 9,595 4 View

Can I concentrate microbial biomass without centrifugation or using membrane filters?

In the lab I work at we do not have many tools for microbiology.

03 April 2014 7,714 1 View

How to confirm the site-directed mutagenesis result without performing NGS?

I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...

08 August 2024 4,645 2 View

I can't see the ssDNA band after performing asymmetric PCR. Is there any way to do this?

After performing symmetric PCR, PCR purification was performed. Afterwards, asymmetric PCR was performed using the PCR purification product as a template, but no ssDNA band was confirmed in the...

08 August 2024 1,668 3 View

How to normalize and take the significance of the MTT OD values with 3 replicates for the same cell-line?

Hi, I have a question about normalizing the MTT OD values for doing the statistical analysis. So, if we have 3 different plates and we call them 3 different replicates, so, first we would...

07 August 2024 8,106 4 View

Why activated CAR-Jurkat cell could not kill targets?

Previously when I co-coluture anti-CD19(FMC63) CAR-Jurkat with Raji with E:T=5:1, Jurkat can eliminate Raji in 24h. However, when I test another CAR construct, although I can dectect totally CD69...

06 August 2024 641 2 View

How much total RNA concentration to be extracted from sorted plasma cells from bone marrow of C57BL/6 mice for RT-PCR ?

i have sorted anti-NP specific plasma cells from bone marrow of C57BL/6 mice at certain times after immunization with variable counts and isolated total RNA using TRIZOL method for RT-PCR using...

05 August 2024 8,835 1 View

Where should the ARS sequence be introduced on a plasmid?

Hi all, I need to introduce an ARS (autonomously replicating sequence) in my plasmid but I'm not sure which position would be the best. Does anyone have any suggestion? A picture of the plasmid...

05 August 2024 1,573 4 View

"A Markov-like Model for Patient Progression"?

A Markov-like Model for Patient Progression" Markov Chain Monte Carlo (MCMC) Markov Chain Monte Carlo (MCMC) is a powerful computational technique used to draw samples from a probability...

05 August 2024 10,079 0 View

Why after performing site directed mutagenesis ,I don't see any colony after transformation?

I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...

05 August 2024 9,059 3 View

Why my colony PCR results of my recombinant bacterial not showing any results?

I am performing ligation of the plasmid and a target gene. The steps I have taken are: 1. Double digestion of the plasmid and target gene 2. Ligation of the plasmid with the target gene 3....

05 August 2024 2,570 3 View

Low-yield gel extraction problem?

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

Laurence Stuart Dawkins-Hall

In essence yes you can
However, for qPCR (not PCR) keep in mind that you can only amplify fragments of 100bp to ensure your primer efficiency is > 90% (ideally >95%)

Md. Shahidul Islam

Agreed with Laurence Stuart Dawkins-Hall

Hend Altaib

I think it is yes. I have tried a similar experiment but, with another pUC and I also agree with Professor Laurence