PCR product purification from gel.

More Doctor Muhandas's questions See All

Can we dissolve plant proteins into human serum?

I want to check the presence of some specific antigenic proteins in a leaf. The system we have is used for detection of proteins using ELISA from serum samples. Is it possible that i crush my leaf...

03 April 2014 859 0 View

Can I use my plasmid as a positive control for ELISA?

I want to perform analysis on some plant samples for the detection of recombinant protein expression. For this purpose I want to use ELISA. Unfortunately I don't have a positive control. Is it...

02 March 2013 7,900 1 View

Storage of leaf sample for recombinant protein

I am working on plant transformation. I want to perform ELISA for my transformed plants and I need to transport my leaf sample to another place. What is the best way to transport the leaf samples...

07 August 2012 3,674 2 View

Multiple bands after PCR using nptII primer.

I am trying to detect putative transformed tomato plants by PCR using Kanamycin specific primer. Instead of getting one band of 720bp( primer designed for) I am getting multiple bands. I optimized...

07 August 2012 1,437 10 View

Southern blotting produced only a strong signal in the control but nothing in the test DNA. Did my transformation fail?

I transformed some plants, grown from initial stages till rooting on 100- 50 mg kanamycin media. After extracting DNA I did PCR using Gene specific primers and got clear sharp bands of desired...

05 June 2012 9,355 5 View

Labeling of southern probe by DIG PCR.

I am trying to label DNA probe by DIG PCR. First I successfully optimized the amplification of gene by simple PCR reagents. Then I used DIG components for PCR reaction mix as recommended by Roche...

04 May 2012 6,285 6 View

Problem in PCR

I designed some primers to amplify a tomato gene. When checked two primer pairs worked very well and gave me required length bands. I repeated this experiment three times and always get bands of...

01 February 2012 1,357 13 View

Purification of PCR product

Hello All, Can anyone have an experience to purify PCR product without using KITs suitable for ligation and cloning. If someone know any manual procedure please guide me about it. Thanks

01 February 2012 5,343 12 View

My question is that is there any way /method to check the function or activity of fruit specific promoter prior to transformation in plant?

I have isolated some fruit specific promoters by PCR amplification. On gel they are giving expected band size. I want to develop a gene cassette in which gene of interest will be under the control...

31 December 2011 1,761 6 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 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

Does anyone have issues using Prepman Ultra reagent for MicroSeq ID bacterial, fungal and yeast sample preparation?

I have been attempting to extract DNA from Bacterial, Fungal and Yeast banked samples (>1e7 cells) using Prepman Ultra reagent and I seem to be struggling to obtain a sequence. Although the...

01 August 2024 2,079 0 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

Dimensions of an MJ Research 96-well alpha module?

Can anyone with an MJ research / BioRad PCR machine from ~2010 or earlier tell me the external measurements (LxWxH) of the removable standard PCR alpha modules that can be removed from a PTC200 or...

30 July 2024 2,867 0 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

Someone have the key or the installer of the program Mx pro 3000 (PCR real time)?

I need to install this program

25 July 2024 4,756 0 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

How do I run a dead well test for a QuantStudio 5 qPCR machine?

I am preparing to run a lot of samples on a 384 well plate and I wanted to test if any well in my PCR machine is dead so as not to lose any data. I would really appreciate help with how I can do...

24 July 2024 8,172 1 View

Falguni Dhwani Desai

You can use the LMP agarose gel method (Sambrook and Russel, Molecular cloning 2001)

Ajay Saini

Hi Dr. Muhandas

Along with the LMP agarose gel...you can also use 'Agarase' enzyme from NEB to digest the matrix and the extraction of DNA band become more easy..

Doctor Muhandas

Thanks Falguni and Ajay for suggestions