What the cloning primers are?

Silvia Caprari @Silvia_Caprari

10 October 2016 0 899 Report

Hi all,

can you explain to me what the cloning primers are? are they also called"diagnostic"? for what purpose are they needed?

Thanks

Badges
Science method

More Silvia Caprari's questions See All

Reference for 0.5 McFarland standard corresponding to 0.08 to 0.13?

Hi, I have a question about the 0.5 McFarland standard. In different websites I read that the for the 0.5 McFarland standard, the absorbance at a wavelength of 625 nm should be 0.08 to 0.13. I...

08 September 2018 4,906 1 View

How do I determine if specific points significantly deviate from the linear regression in a scatter plot?

Hi everyone, I would like to know how I can determine if a point significantly deviates from the linear regression when I plot the data in a scatter plot. I am using excel and I made a scatter...

06 July 2018 5,349 8 View

How to measure density with ImageJ in images that have different magnification?

Hi all, i am new in image processing and using ImageJ. I want to measure the density in areas of pictures that were originally taken at a different magnification. I attached an example that I hope...

05 June 2018 7,340 4 View

Theoretical question about western blot?

Hi, I am new with Western Blot and I need a few clarifications about the use of blocking and incubation solution. I understood that blocking is done to saturate the membrane with proteins...

11 December 2017 4,469 4 View

Preparation of sodium phosphate buffer?

Hi, I am preparing a sodium phosphate buffer solution at 0.1 M pH 7. For the preparation I am following this instruction that requires to mix 1 M Na2HPO4 and 1 M NaH2PO4 like this: 57.7 ml 1 M...

10 November 2017 6,602 3 View

How to calculate the enzyme activity from absorbance?

Hi, I have a question about how to calculate the enzyme activity from absorbance. I have absorbance values for different concentrations of my samples at different times. As far as I understood, I...

08 September 2017 1,836 4 View

Can you help me with understanding how to determine the beta lactamase activity?

Hi all, I would like to ask you a question about the data analysis in an enzymatic activity assay. It is the first time I deal with this kind of stuff. I have a kit for the beta lactamase...

08 September 2017 1,279 5 View

Which statistical test is most appropriate for my experiment?

Hi, I need some suggestions on which statistical tests are most appropriate for the analysis of my experiment. Basically I have plates with different antibiotic concentrations in the medium. The...

08 September 2017 8,251 15 View

Cutting bands out of the agarose gel?

Hi all, I have quite a stupid question about safety precautions when cutting out bands from an agarose gel under a UV transilluminator. I am aware that you need to wear lab coat, long sleeved...

05 June 2017 3,145 1 View

Don't I loose the plasmid with a new eluition in water during the miniprep?

Hi all, I made different minipreps starting from the same sample of bacterial culture because my intention was to increase the concentration of plasmid in my final plasmid preparation. Now I have...

05 June 2017 3,071 4 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

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

Anyone having idea about VN primer for miRNA primer design ?

How to design VN primer to attach with universal reverse primer

05 August 2024 2,116 3 View

How can we now if the promotor in the vector is well?

Hi, everyone. I want to transfer two genes in to the pseudomonas bacteria that isolated from the soil. The bacteria that I want to use for cloning haven't been identified and not be sequenced,...

02 August 2024 3,987 1 View

I'm optimizing a tetra-ARMS PCR protocol with amplicon sizes: 165bp, 123bp and 93bp. Why, in the gel, only 165bp is visible while I'm expecting all 3?

It's an end-point PCR protocol. I'm using 1.5% agarose gel with SyBR Safe dye and TBE as a running buffer, visualization on BioRad XR+ system. I was primarily thinking of primer efficiency,...

01 August 2024 4,673 4 View

PCR showing no bands - Master mix and primers didn't mix?

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

How to retain amplicon yield after Ampure bead cleanup, following a 2-stage PCR protocol?

Hello all, I have been trying to follow a 2-stage PCR protocol used to amplify barcodes of a large yeast library, as per Nyugen et al. (2022) -...

30 July 2024 841 2 View

Do you mix and add all primers together in a singel PCR reaction (4 total primers for 2 mutations) when using Agilent's multi-site mutagenesis kit?

I want to introduce 2 mutations using Agilent's multi-site mutagenesis kit, I designed the primers using their online tool and it gave me 2 sets for primers (1 set for each mutation) so I was...

25 July 2024 6,517 3 View

Illustra™ MicroSpin™ G-25 columns what it is used for?

Hello, We found three packages of Illustra™ MicroSpin™ G-25 columns in the cabinet of an unused lab. They are very old but have never been opened. I have never used this kit before, and I couldn't...

25 July 2024 4,927 3 View