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

What is the difference between PCR and gradient PCR?

Any body can explain about the PCR and gradient PCR technology using an example?

11 December 2017 4,964 2 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 can I represent SDS-PAGE results for machine learning analysis?

I am trying to classify and analyze the results of an SDS-PAGE based array for bacterial detection using machine learning, but I have trouble finding the best way to represent the results with...

27 February 2021 9,176 3 View

Why my agarose gel staining sometimes looks not homogenic but like gradient (see the figure)?

I am worried about this overexposure of the upper part of the gel in the picture. Is it possible that this is the result of too much concentration of ethidium bromide? Why is there such a big...

25 February 2021 8,140 3 View

EMSA troubleshooting. Protein-DNA complex not migrating, cold probe lane with multiple bands. Can anyone help?

I'm running an EMSA to show the DNA binding activity of recombinant proteins versus the Wild type. Previous assays run by my research group using a different test system show the protein-DNA...

24 February 2021 8,325 1 View

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View

How to manage western blot for proteomics data validation?

Hi Everyone, I am a bigginer in western blotting and I am trying to use it for validation of my proteomics data. I have: two regulated proteins with molecular mass of 40 and 73 kDa one control...

23 February 2021 6,671 4 View

Alginate and silk fibroin hydrogels toxic to cells?

Hello, I asm currently working with alginate and silk fibroin hydrogels. I prepared a blend of alginate and aqueous silk fibroin (straight from dialysis) and resorted to ionic crosslinking to...

23 February 2021 8,249 2 View

Diltuions for a Western blot?

I need help using the equation C1V1=C2V2 figuring out how much antibody to add to use for a western blot. The stock was origianlly 3.45 ng/ul but diluted 1:10 so now its 0.345 ng/ul. I need to run...

22 February 2021 9,901 3 View

Why the nucleic acid gel melted after running for 30min?

Hi all, I proceed with nucleic acid gel electrophoresis recently several times, but a wried phenomenon is they always melted after running over 30min. My gel concentration is 1% percentage...

22 February 2021 9,896 3 View

Sanger Sequencing results not matching experimental results of Restriction Digestion, colony PCR and protein expression?

I have cloned two domains of a gene into two bacterial vectors using cut and paste cloning with different RE sites. I checked the putative clones by colony PCR and Restriction Digestion (fallouts...

18 February 2021 6,736 4 View

How could the concentration from RNA extraction is really low?

Hello everyone, currently, I'm trying to do RNA extraction from seeds of Arabidopsis. I'm using the protocol below: RNA isolation from siliques, dry seeds, and other tissues of... I use about...

17 February 2021 7,583 4 View