PCR product purification from gel.

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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 783 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 7,823 1 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 1,327 10 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 3,595 2 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 9,278 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 6,202 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 1,273 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 5,265 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 1,666 6 View

How do I increase the yield of my PCR product?

Hello, We would like to increase the yield of our PCR product. We are running a series of PCR reactions that is targeting ~1.1kb sequence. We begin each reaction with ~400pg of template DNA...

02 March 2021 4,029 3 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

How to address this issue in the analysis of Real-Time PCR results?

To dear Researchers, I was analyzing a series of concentration for estimation of Real-Time PCR efficiency. The concentration was 1:10. I used MS-excel to evaluate Slope. The result of slope was -8...

01 March 2021 8,683 4 View

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Does anyone have the experience of using Taq Man probes in the QIAGEN Rotar- Gene qPCR machine?

01 March 2021 5,311 1 View

MiRNA qpcr problem?

Dear All, mirna primer showing some problem in the melting curve? any idea why? As attached is the melting curve. The forward sequence is obtained from miRBase and reverse primer is universal.

28 February 2021 5,008 4 View

Runs of nucleotides in Sanger sequencing?

I performed site directed mutagenesis, transformation, and then I sent out plasmids for Sanger sequencing and found out that there is extension of DNA just before the stop codon. I am not sure...

27 February 2021 547 3 View

Can thermal cycler machines be problematic during PCR?

Hi, my question is about the heating of thermal cycler machines and I hope some of you had experienced a similar thing previously. There are two thermal cycler machines in the lab(BioRad) and for...

26 February 2021 4,777 4 View

Calculating volume of DNA required from DNA concentration?

Hi I am a bit confused. They are asking me to find out the volume of DNA required in ul (a total of 30-100 ng for genomic DNA) from the DNA concentration in the nanodrop reading which was 404.8...

26 February 2021 5,029 2 View

Has anyone used the Illumina ITS1 primer sets?

Hi everyone, Illumina provides a list of primers to amplify with high taxonomic coverage the ITS1 region for further fungal sequencing, but I cannot find the exact amount necessary of each...

25 February 2021 6,969 3 View

Why do the PCR bands from the cDNA templates (extraction from the cell after transfection) show different size from the band of the plasmid template?

After transfection with the plasmid ( linearized ) and subcloning of the cell lines, RNA was extracted from the cell and then reverse transcripted to cDNA. When PCR reactions were run to verify...

25 February 2021 5,712 3 View