Do forward and reverse primers hybridize as a whole quantity, or some of their complementary nucleotides to the DNA template?

02 March 2015 851 0 View

What is the formula to calculate Tm of primers F: TCAAAGGAATGTTGGG and R: ACGCACATAGGGATTG to use in PCR?

02 March 2015 9,414 8 View

Is anyone familiar with the Ym30 pair primers?

09 October 2014 5,359 1 View

What is the role of MgCl2 in PCR?

PCR amplification effects by the MgCl2 concentration. That what happen in an experiments I carried out. I need more advices and recommendation until what concentration the amplification could...

08 September 2014 7,192 12 View

I have a problem with tailed primers. The forward primer is cut and only M13 left. Can anyone help?

I am using M13 tailed primers with the forward which initially starts the amplification PCR reaction. When the part of the Forward Primer is cut off, can the M13 tailed primer amplify later on...

08 September 2014 2,533 3 View

Where can I get Malayisan Yam samples?

I am student at Nottingham University, working on a project on Yam in Malaysia. I am having difficulty finding where to collect samples, because I am an international student. I would be very...

04 May 2014 8,862 1 View

What are some methods for DNA Pooling (grouping)?

When putting together more than one sample of DNA in order to run the PCR gradient, what are the methods to group different samples of same species of DNA together besides having the Nano Drop...

04 May 2014 2,603 1 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

Unable to detect a truncated protein variant, but I can observe a truncated cDNA from Sanger?

Hi, so basically... I am trying to mimic a clinically observed condition wherein a frame-shift mutation happens only at one allele of the patients, giving rise to a truncated variant which evades...

17 February 2021 1,130 6 View