B R Siddharth

30 Questions 16 Answers 0 Followers

Questions related from B R Siddharth

I have a 11kb fragment and I am trying to do IVT but every time I perform IVT a smear like band observe also it doesn't match the size any solution?

I am using 1% Agarose gel for running RNA band and the kit I am using is T7 NEB IVT kit I have also used RNase Inhibitor but nothing changed.

15 March 2025 3,226 2 View

Why do I have poor DNA quality and yield after gel extraction and clean up ? i want to perform IVT and do electroporation any suggestions?

I am working on my cloned fragments in a low copy number vector so generally, i get low yield of my recombinant plasmids. I have two different genes in two low copy plasmids i want to perform...

27 February 2025 8,284 5 View

Cloning of flaviviruses is difficult i have noticed that cells loose their transformation ability after cloning fragments inside vector any advice?

As flaviviruses genome is very frequently prone to mutation hence it is very difficult to clone whole genome inside vector what kind of vector and cells can sustain such amount of cloning...

12 July 2024 4,484 0 View

I am not getting proper colonies while ligating one of fragments in my vector the colonies seems to be normal but in light it seems lysed any reason?

I am working on cloning of a gene fragment inside my vector everything we tried to do correctly like making insert and vector with utmost care but after ligation the morphology of colonies seems...

12 July 2024 9,206 3 View

Why generally we prefer phenol chloroform extraction method for DNA purification as i always face difficulty in getting good concentration why?

phenol chloroform extraction method is used for purification of nucleic acids DNA/RNA

04 April 2024 8,311 0 View

After phenol chloroform extraction the nucleic acid concentration is around 20ng/ul but after loading 1ul/2ul its still not visible why?

phenol chloroform extraction method is used to purify nucleic acids to obtain a good concentration.

04 April 2024 7,286 0 View

Can anyone suggest some specific bacterial cell line for cloning viruses, specifically flaviviruses i have used cell like TOP-10/DH10B/DH5 suggest?

as viruses are notorious for being prone to mutation at high rates its very difficult to clone them though general cloning procedures and cells so in order to solve this problem need specific cell...

04 April 2024 9,072 5 View

I have designed primers for side directed mutagenesis for bac plasmid for deletion of Kpn-I site and transformed it in DH5 cell but no colonies why?

BAC vectors are use for cloning purpose and are good for cloning large fragments though they have low copy numbers.

26 May 2023 5,171 2 View

How do we decide what primers do we have to take in cloning during reverse genetics platform experiments please discuss some necessary tools?

reverse genetics platform is used for cloning platform where cDNA is made from Viral RNA and is used for cloning

15 February 2023 4,676 0 View

Can we proceed in cloning without isolation of RNA from viruses is there any alternate strategy to do so ?

durring reverse genetics platform RNA isolation is a necessary step and from there we proceed for cloning

15 February 2023 5,511 2 View

Are competent cell transformed with ligation product more resistant to transformation than normal plasmid transformation if so how to increase it ?

transformation after ligation is nessesary step to get desired product in large amount.

04 February 2023 2,931 4 View

Can centrifugation at 11000g after transformation in E.coli DH5 alpha cells will result in lesser or no colonies ? what is the ideal RPM or G ?

after bacterial transformation bacteria need to be speeded on a percitiplate to do so we need to pallet bacterial cell and it is subjected to centrifugation.

03 February 2023 2,861 8 View

When i am thansforming ligation product inside DH5 alpha cells now i am not getting even as ingle colony what coulb be the possible resons for this?

for construction of particular clone logation is a nessasary step and in order to do so ligation is required and sometimes colonies are not obtained.

24 January 2023 7,457 0 View

After transforming the ligation product on LB-amp plates colonies are obtained but why there is no growth after inoculations for 16 hrs in 2XYT-amp ?

cloning and trasnformation of bacteria containnig ampicilin resistant gene

16 January 2023 5,767 4 View

What is the easy way to make 6x loading dye?

loading dye to track DNA smaples on electrophoresis and load them on wells

07 October 2022 8,058 3 View

How to remove unwanted noise from uv-vis spectra? any suggestions regarding nano particles synthesis to get a sharp spr?

for synthesis of nanoparticles, a sharp peak must be obtained around 410-450nm

10 December 2019 5,436 5 View

Any idea about these chromosomal abnormalities i would like to know their names?

chromosomal abnormalities induced due to biological silver nano particles in plants

03 October 2019 4,377 0 View

Any suggestions regarding the graph obtained for nano-particle synthesis how we can improve this graph to obtain SPR ?

We have attempted to synthesize Silver (SNPs) from Fenugreek any suggestions regarding improvement and how to interpenetrate this graphs we have used 3mM Silver nitrate and 30% plant extract.

25 September 2019 9,834 1 View

What is the absorbance peak or SPR for nanoparticles in uv-vis spectroscopy?

Surface plasmon resonance of silver nano particles synthesis

25 September 2019 5,026 5 View

Need suggestions on evaluation of different types of chromosomal aberrations induced mitosis root tips due to genotoxicity article or references ?

root tips are used to evaluate genotoxicity caused due to different agents like silver nano and industrial effluents.

23 August 2019 9,188 0 View

How to calculate size of silver nano particles from UV-VIS spectrometer data having SPR?

SPR indicates formation of silver nao in UV-vis spectrophotometer data it gives SPR Peak

19 August 2019 6,048 4 View

What is the reason for this type of graph in 300-700nm range for silver nanoparticles synthesis?

Uv- vis spectro reading in 300-700nm range for silver nano sunthesis

16 July 2019 2,022 14 View

Suggest a good method to study chromosomal aberrations in plants how to prepare its slides for microscopic observations?

to study effects of silver nano particles on plant Genotoxicity

13 June 2019 306 1 View

How one can study Chromosome aberrations in plants as a monitoring system using silver nano particles what are its applications?

silver nano particles create chromosomal aberrations in plants

10 June 2019 3,609 2 View

What is surface Plasmon resonance durring silver naoparticle synthesized from plant extract?

silvernano particles are produed from palnt extract various chemical and physical process can be used to produce silver nano particles

05 June 2019 7,012 5 View

What are microgreens and what is its sinificance as a functional food ? what is importance of pak choi verities how these are grown under LED lights?

microgrens are used as a food source from plants

05 June 2019 2,030 2 View

If you were a botanist that was going to the Moon or Mars as an astronaut, what kind of research project would you develop?

This is an open thought question for future xenobotanists. I read that NASA is hiring a new round of astronauts for their Mars project, and it made me think about botany that is not on...

01 January 1970 3,134 3 View

how one can optimize the synthesis of AgNPs from plant extracts

optimization of Ag NP synthesis

01 January 1970 7,805 1 View

What is C-Meta phase why it occurs during genotoxicity?what is sticky meta phase anaphase with ring formation?

genotoxicity at chromosomal level

01 January 1970 8,731 0 View

What is the diffrence between characterization of NPs by DLS and SEM?

Nano particle characterization

01 January 1970 5,552 6 View