Is it necessary to purify PCR products before cycle sequencing?

More Saptamita Goswami's questions See All

Could you please advise on the optimal amount of exopolysaccharide to extract from a 500 ml bacterial culture?

12 December 2023 9,480 0 View

Why does a cyclic voltammogram have multiple anodic or cathodic peaks for certain materials?

I took the CV of nickel ferrite using carbon black as the counter electrode, the voltammogram shows multiple reduction peaks. why does that happen?

21 September 2023 9,475 1 View

Why does the CV scan taken using different instruments show different results?

The CV plots of buffer solutions having PH 3-10 was done using CH instrument as well as using AutoLab. Both the results have different peak positions and different shapes.

21 September 2023 1,038 1 View

What kind of contamination is this?

I am working with U87MG cell line and I got contamination in one of the flasks. Can anyone tell me what type of contamination is this?

21 September 2023 8,669 0 View

How to identify alpha and beta cellulose on the basis of extraction method?

In my study we used chemical treatment for the extraction of cellulose.

17 August 2023 4,264 1 View

Free softwares for adsorption study its kinetics isotherm and thermodynamic and also which software is used for error data analysis?

In my study i am using nanocellulose based membrane for the adsorption study of dye and heavy metals

26 July 2023 2,307 4 View

Is there any postdoctoral position in foreign universities (outside India)?

Dear fellow researchers and professors, I am currently pursuing my PhD in the area of industrial management, and looking for postdoc in foreign universities outside India. If you are aware of some...

23 June 2023 457 0 View

What are the Challenges and Opportunities in Advancing Adult Education in India?

India has made progress in its literacy rate because of adult education programmes but unfortunately adult education in India is seen as a discipline for literacy only. This discussion will...

04 May 2023 6,940 2 View

What is the range for MLVSS in anaerobic reactors?

typically in an anaerobic filtration system or anaerobic reactor treating high strength wastewater

06 April 2023 5,988 1 View

Measurement of Reactive oxygen species (ROS) in Blood using DCFDA dye ?

I want to measure reactive oxygen species in whole blood. Can anyone please suggest a protocol or method for it using 2',7'-dichlorodihydrofluorescein diacetate (DCFDA) dye ?

09 February 2023 3,599 1 View

Can I base on reverse DNA sequences to perform alignment, convert to amino acids and GenBank submission?

I have reverse sequences (AB1 format), can I base on reverse DNA sequences to perform nucleotide alignment, convert nucleotides to amino acids and deposit the sequence in GenBank database?

11 August 2024 5,138 1 View

Do interactions between biosphere, carbon cycle, & water cycle impact global warming & interaction between atmosphere & hydrosphere?

How do interactions between the biosphere, the carbon cycle, and the water cycle impact global warming and interaction between the atmosphere and the hydrosphere?

09 August 2024 3,291 2 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

"A Markov-like Model for Patient Progression"?

A Markov-like Model for Patient Progression" Markov Chain Monte Carlo (MCMC) Markov Chain Monte Carlo (MCMC) is a powerful computational technique used to draw samples from a probability...

05 August 2024 10,079 0 View

Why after performing site directed mutagenesis ,I don't see any colony after transformation?

I want to introduce a point mutation (change in one nucleotide) into my gene of interest (DNA binding domain) I have designed primers as recommended on the Data sheet of the kit : -Both primers...

05 August 2024 9,059 3 View

Why my colony PCR results of my recombinant bacterial not showing any results?

I am performing ligation of the plasmid and a target gene. The steps I have taken are: 1. Double digestion of the plasmid and target gene 2. Ligation of the plasmid with the target gene 3....

05 August 2024 2,570 3 View

Low-yield gel extraction problem?

I am having an issue with my gel image where my PCR product is not appearing very bright on the gel. When I perform gel extraction, the A260/280 purity value is very low. I used the Qiagen gel...

05 August 2024 9,798 3 View

Identify the eight key processes of excellent supply chain management and discuss how each of these processes impacts the end customer?

Read the journal article by Douglas M. Lambert, “The Eight Essential Supply Chain Management Processes,” Supply Chain Management Review, Vol. 8, No. 6 (2004), pp. 18-26

04 August 2024 9,919 4 View

Waleed Mohammed

https://pdfs.semanticscholar.org/d1ae/cec02e3586c044eb11ae6779e3baf616d244.pdf

Giorgia Pertile

https://www.edgebio.com/products/performa%C2%AE-spin-columns-13266

We use this before sequencing plus cleaning with ExoSap.

Sibnarayan Datta

Yes it is necessary, if you want to have a good sequence data.

best wishes,

Katie A Burnette

I've had better luck cleaning up reactions with "Exo-SAPit" reagent. It removes unused primers and dTNPs. Of course, most sequencing facilities will offer to clean it for you - look at the cost and decide what's best for your lab.