140 Kb Electrotransformation in E.coli. How should I do it?

More Victoria Nugnes's questions See All

Is this a mixed methods approach?

For my PhD dissertation: If Chapter 1 is a systematic literature review (QUANTS/QUALS) while Chapters 2, 3, and 4 are empirical chapters (QUALS), is this considered a mixed methods research...

16 June 2024 3,220 5 View

In the case of Collagen Cell Carrier® membrane 34 mm for 6 well cell culture plate (Viscofan), can you suggest a paraffin embedding protocol?

I am intrested in the preparation of collagen carriers, dehydration, paraffin infiltration, and paraffin embedding. Thanks in advance.

05 June 2024 3,831 0 View

How can I detect a small amount of Protein without overexpression?

Hi, are there other methods beside Western Blotting to detect a specific protein? I‘ve isolated mitochondria and want to show that hPar17/Pin4 is present in mitochondria. The Western Blots didn’t...

24 May 2024 5,500 3 View

How many keratinocytes and fibroblasts are contained in 1 mm^2 of skin?

Good afternoon, everyone. Perhaps someone knows how many keratinocytes and fibroblasts are contained in 1 mm^2 of skin. Or the ratio of these cell types may be known. If there is a link to an...

15 May 2024 5,256 0 View

Which antibodies can be used to detect mitochondria in western blots?

Hi! I´m currently trying to isolate Mitochondria from HCM cells. For my western blots I need an additional antibody against a mitochondrial protein which is bigger or smaller than 14/17 kDa. I was...

24 April 2024 8,814 2 View

How do I block mouse Fc receptors when my primary antibody is raised in rat?

Hi, I have having an issue with the Fc blocking stages when carrying out flow cytometry on a murine macrophage cell line. I am using a rat anti-mouse antibody against the scavenger receptor...

22 April 2024 3,466 2 View

Thematic analysis or grounded theory?

i am currently conducting a critical appraisal of a research article and the data analysis is explained as: interviews transcribed verbatim and participants were given the option to review and...

21 April 2024 1,821 7 View

Does anyone have ultrasonic audio files of male mice?

For a project I'm included in I'm desperately in need of audio recordings of male mice. They need to be original files recorded with the correct equipment. I need them for research on gait...

19 March 2024 4,959 1 View

How lower year interacts with higher year students in America?

lower year students attitude towards higher year students in America

09 March 2024 4,747 1 View

What is my research gate id?

29 November 2023 3,950 0 View

Can I base on reverse DNA sequences to perform alignment, convert to amino acids and GenBank submission?

I have reverse sequences (AB1 format), can I base on reverse DNA sequences to perform nucleotide alignment, convert nucleotides to amino acids and deposit the sequence in GenBank database?

11 August 2024 5,138 1 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 can I apply boundary conditions in an orthotropic steel deck numerical model using ABAQUS software?

I am trying to simulate vehicular loading on an orthotopic steel deck bridge section in ABAQUS software. The red arrow mark in the attached figure indicates the direction in which the vehicle will...

08 August 2024 719 0 View

Does crude extraction using NaOH and Tris work well with Fungi?

I'm trying to find a DNA extraction method for fungi that does not require equipment and heating. Is there anyone who can suggest an alternative option? Thank you

08 August 2024 4,733 2 View

Is there any way to quantify bacterial and fungal cells in their mixed culture?

I am working in fungal fermentation of soybean meal and there is bacterial growth in them at times. I am trying to quantify fungal cell counts and bacterial cells; but I haven't been able to do at...

07 August 2024 7,535 4 View

Unusual intensity drop in some sections of chromatograms in DDA?

Hi, we have measured tryptic peptides using both DDA and DIA method on QExactive. In DDA replicates i saw unusual intensity drops occurring at the same sections of chromatograms in DDA replicates...

07 August 2024 3,218 4 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

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

Ibrahim Bitar

Did you try One Shot™?

Did you try One Shot kit ?

One Shot kit is one of the best kits that deals with transformation. give it a try

One Shot kit is one of the best kits that deals with transformation. give it a try.

Victoria Nugnes

@ Ibrahim Bitar I am looking for electrocompetents since i ve read that are muy efficient for large DNA because with an electrick shock the pores are bigger