For cloning of large PCR product (20 kb) Any recommendation ?

More Suppiah Paramalingam Sivalingam's questions See All

What are the factors for the failure of livestock-integrated and segregated farms?

When dairy and livestock production fails in the country, it can have significant economic impacts at various levels, including the local, national, and even global economy such as Loss of Income...

23 February 2024 1,736 0 View

Until which passage is cells considered as Primary cell?

I am currently working with human lung epithelial and fibroblast. I would like to know which passage is best for invitro studies as well. Kindly advise. Thank you

02 October 2023 6,517 5 View

CDNA synthesis or qPCR did not work?

Hi everyone! I extracted RNA from urine pellet using phenol-chloroform method. The yield of RNA was 160ng/uL. 3uL of RNA was used for cDNA synthesis in 20 uL reaction using Go-Script kit....

05 February 2021 6,874 4 View

How to improve the techniques for gel picture of RNA?

Dear all, I have extracted RNA from urine samples using the Phenol-Chloroform extraction method and obtained the yield (~83ng/uL) on Quantifluor Fluorometer. But I could not get a clear band on...

13 December 2020 6,740 3 View

In TGA analysis weight loss % graph after 800 degree it is going upwards?

Kindly help me to analysis my TGA weight loss % graph up to to 800 degree weight loss is decreases whereas after 800 degree weight loss increases. can some help me to understand. Thanks in advance

21 March 2020 8,060 9 View

How to remove organic molecules selectively in top surface?

We are preparing organic molecules functionalized ZnO nanords (vertically aligned). For my application, I don’t need organic molecules functionalization on top surface of the ZnO nanorods. How to...

22 February 2020 2,138 2 View

What is the right statistical test?

Hi , I am doing a within group experiment. I have one independent variable with three levels and one dependent variable with three levels. What is the right statistical test for this experiment....

06 April 2018 8,568 6 View

What is the concept of increasing convective heat transfer coefficient with increase in Renolds number?

Generally Heat transfer coefficient of fluid increases with the increase in Reynolds number or velocity of fluid when it flows through a pipe. What parameters of fluid increases that causes an...

10 February 2017 6,635 8 View

Which is a better fixative for Focus forming assay..ice cold methanol or paraformaldehyde?

Anyone have done comparison of ffu assay using ice cold methanol and paraformaldehyde? which is better in terms of getting a clear foci? Any tips to get a better number of foci?

16 November 2016 3,818 3 View

What are the scope and features of 2D transition metal dichalcogenides, to prepare by wet milling and used for energy related applications?

Graphene is not optimistic in semiconductor industry, on the other hand other 2D materials have lot of scope in it. I guess we cannot make defect free and lengthier 2D materials using wet milling....

03 March 2014 9,205 3 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

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

Does crude extraction using NaOH and Tris work well with Fungi?

I'm trying to find a DNA extraction method for fungi that does not require equipment and heating. Is there anyone who can suggest an alternative option? Thank you

08 August 2024 4,733 2 View

Are there instances where molecules with larger molecular weights exhibit greater mobility than those with smaller molecular weights?

Hi, I know that low molecular weight (MW) molecules generally tend to have higher mobility, while high molecular weight molecules tend to have lower mobility. However, in my experimental...

06 August 2024 1,495 2 View

Weak DAPI staining after immunohistochemistry - how to improve?

After immunohistochemistry of previously fixed in PFA and EtOH and then frozen 20 μm sections of zebrafish brain, DAPI staining is very weak (right) compared to the same sections stained without...

05 August 2024 9,637 2 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

The Curse of Evolution and Complexity?

Brain and body mass together are positively correlated with lifespan (Hofman 1993). The duration of neural development is one of the best predictors of brain size, and conception is the best...

05 August 2024 6,247 3 View

For an in-vitro drug release study, what molecular weight cut-off (MWCO) dialysis bag is required for a 117 kDa protein?

kindly reply me. Thanking you in advance.

05 August 2024 7,727 4 View

How to isolate lymphocytes from mouse spleen?

I have tried several times to isolate lymphocytes from mouse spleen, but all attempts have been unsuccessful. I tried most available protocols. I used different dissociation media (HBSS with Ca...

04 August 2024 9,913 7 View