When i purified my digested plasmid from the agrose gel ;and try to run of the gel again it just float from the well.Is my plasmid is contaminated ?

01 January 2018 0 4K Report

is my plasmid is contaminated with ethanol;how to avoid this contaminated

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I want to do protein expression in E. faecalis JH22 by using anhydrotetracycline (ATc) to induce protein expression .Need protocol for expression?

I want to use E. faecalis JH22 as competent cells to express my protein using anhydrotetracycline (ATc) to induce protein expression i need some standard protocol if someone has use it before.

06 July 2019 9,595 2 View

How to check the genome sequence of locus A of Enterococcus feacium TX16 species;any site for this ?

website,genome sequence

01 February 2019 7,240 2 View

I want to use PET-Duet vector but i donot protocol how to do restriction cutting with two enzymes ;if i want to insert two gene of interest?

As PET duet vector is for cloning of two gene of interest;i want the protocol for it.

06 July 2018 9,166 0 View

Why are my gel electrophroesis samples are not moving?

I run gel electrophoresis to confirm digestion of my plasmid however my samples are remaining at the start of the gel. I have remade wash buffers and carrier buffers but am still not seeing my...

31 December 2017 6,316 0 View

POPINF plasmid i want to send for sequencing i donot what type of primer i have to use company have standard primers?

can anyone please recomend me any primer

10 November 2017 633 1 View

Ammonium sulphate precipitation method for protein , after centrifugation when i got the pellet,in which buffer dialysis of pellet has done to NMR?

in which i resuspend my pellet against which buffer i have to dialysis.

10 November 2017 5,654 2 View

I want to express my TX6119 strain ;i run it as control without injecting IPTG i saw no band?

Tx6119( M15 containing fragment of SwpA26-226 from TX82 cloned into pQE30),when i run it as control without injecting IPTG i never saw any band on SDS-gel.

09 October 2017 3,269 4 View

I am using Sybre Safe to stain the gel electropheresis;i 5microlitre in each welll but donot got any band, i just saw ladder band ,i am adding 6x dy?

Can anyone suggest why i am not getting any bands

09 October 2017 5,763 3 View

How much i add amipicillin and kanamycin into pQE30 (Qiagen) contains cloned fragment and transformed into M15 ( (pREP4) cells to do miniprep?

TX6119- Fragments were cloned into the expressionvector pQE30 (Qiagen) and transformed into M15 (pREP4) cells(Qiagen).i want to make culture in LB broth to do miniprep i am confuse how much i add...

07 August 2017 401 4 View

What type of positive and negative control;can be use for Histag antibody in Western blot?

Positive and negative control

05 June 2017 6,000 3 View

Does anyone know what might be causing the smeared bands on my western blot?

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

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

Why is my ASO degraded quickly on agarose gel?

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

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

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

Recovery rate after peptide desalting using Sep-pack C18 1 cc Vac Cartridge?

Recently I'm trying to use Sep-pack C18 1 cc Vac Cartridge (Waters: SKU: WAT054955) to desalt digested peptides. However, the recovery rate is usually 40-60%. What is the normal recovery rate?

31 July 2024 6,544 11 View

Transfection in HEK293T cells?

Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...

31 July 2024 9,892 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