Why I got non specific band after PCR ?

I did PCR reaction without using any PCR kit . The result is not good enough. I got heavy low non specific band and probably primer dimer (picture attached). First, I did with annealing...

09 October 2019 1,794 4 View

Why I failed to get good band after plasmid isolation?

I tried to isolate plasmid DNA from Agrobacterium tumefaciens using both manually and using kit and performed the electrophoresis on 0.5%, 0.8% and 1.0% agarose gel. The ladder showed the...

08 September 2019 2,097 2 View

Can you identify?

After culture the sample on PCA I got the colony like this. Can anyone identify upto species? the plate 4 is serial diluted.

10 November 2018 4,576 0 View

How can I solve primer dimer problem without changing primer?

For primer dimer there are many reasons. One is primer designing. Is there any way to solve the problem with the same primer that have been designed?

05 June 2018 3,619 2 View

How this can be interpreted?

After PCR followed by gel run every time I got this picture. Can anybody tell me why this happened and what will be the solution? In these two pictures, one gel had been run after putting 1kb...

04 May 2018 5,260 0 View

Any conferences in Biotechnology before Feb 2018?

09 October 2017 2,468 2 View

What does it mean by primer length 20mer?

I designed some primer for my experiment and I do not know the actual length of the primers which I must need to know for getting the result. I got a manual with the primers supplied by the...

09 October 2017 1,948 4 View

Preparation of water extract molecule

While I am preparing WEB (Water extract molecule from Streptococcus pneumoniae) after 6 hours shaking it supposed to centrifuge in 10000rpm. But my centrifuge machine was shown error that is why I...

01 January 1970 2,828 2 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

Research on Supply Chain Management in Indian context?

Hi, I am planning to apply for the PhD degree in the Supply Chain Mgt. with specific area of "Cold Storage warehouses" during Pandemics and wars. Where lock downs and shut downs are frequent....

02 March 2021 285 2 View

PACYC PCR not working after several attempts. Would anyone have any suggestions?

So, I have been trying to run a pACYC PCR which will be used later on for a Gibson Assembly. However the PCR is not working. I have already tried gradient PCR and changing extension time; however...

02 March 2021 1,146 2 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

No title

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