Is it necessary to exclude stop codon in the primers sequence synthesized for clonning a gene into pET vectors with C-terminal His-tag?

More Parismita Kalita's questions See All

Under what conditions absorption and excitation spectra of a same species can be different?

I took absorption and excitation spectra of a same species in the same solvent. Ideally I should get the excitation spectra almost similar to the absorbance spectra. But I got the excitation...

13 December 2020 8,965 2 View

Is it necessary that during peer review process the revised version of the manuscript is reviewed by the same reviewers who have reviewed it first?

During peer review process, as we all know we modify our articles based on the reviewers comment that we get. But when we submit our revised version of the manuscript, is it necessary that the...

29 June 2020 8,111 7 View

How do journal editors approach when multi reviewers submit conflicting decisions?

Suppose a paper have been reviewed by three different reviewer's. Two reviewer's suggested acceptance of the article, and one suggests not to accept the article. In such situations, what the...

02 May 2020 4,918 7 View

What are the Human Resource Practices of Indian Oil Corporation Limited (IOCL)?

I am researching for better Human Resource Practices which may be universally accepted by all organisations. Request all to share the answers/ links/ any documents/ videos

14 April 2019 6,418 3 View

How to perform SPAM steganalysis ?

I have downloaded the file from below link . http://dde.binghamton.edu/download/spam/ Thank you in advance.

17 September 2018 4,827 6 View

How does the electron mean free path or electron-lattice mean free path depend on temperature?

Mathematically, how does the electron mean free path or electron-lattice mean free path depend on temperature? Is there any formula linking the two?

12 December 2017 2,208 2 View

What will happen if we inject streptozotocin before nicotamide to induce diabetes in rats?

In general we induced nicotamide first which will be followed by streptozotocin after 15 mins to induced diabetes in rats. but i would like to know if we reverse the process. what about their...

09 December 2016 9,154 9 View

What should be the concentration of citrate buffer for anti-diabetic study in rat?

I will induce drug(Streptozotacin-Nicotamide) on rat as per the body weight. i want to know the concentration(molarity/normalioty) of citrate buffer ph-4.5, so that i can mix the drug and induce.

28 November 2016 8,240 3 View

Is there any method to compare two rank list and find out the better one?

Let's say I have two ranking list of homogeneous items. The rank list is prepared based on two different parameters, to find out the best items. Now is there any methodology that is helpful to do...

17 August 2016 5,420 3 View

What is the maximum concentration of imidazole that can be used to elute strongly bound protein from Ni-NTA column?

Lots of protein remains bound to the matrix even after elution with buffer containing 500mM Imidazole.

08 June 2016 4,897 2 View

Can linear DNA be used as template for site-directed mutagenesis?

Is it possible to induce site-directed substitution mutation by quick-change method on linear dsDNA? or it has to be cloned in some vector? If yes, should it be treated with the Dpn1 enzyme...

03 March 2021 401 4 View

In PRIMER, is it possible to downweight certain taxa in multivariate community datasets to reflect uncertainty around identification??

I have a dataset with about 80 different species. As usual, some species are very easy to identify with certainty whereas others are more difficult, which means that I am less certain of my...

03 March 2021 8,066 4 View

PACYC PCR not working after several attempts. Would anyone have any suggestions?

So, I have been trying to run a pACYC PCR which will be used later on for a Gibson Assembly. However the PCR is not working. I have already tried gradient PCR and changing extension time; however...

02 March 2021 1,146 2 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 View

How to calculate the annealing temperature of a primer pair?

I have to amplify a gene and my primers just reached. The Tm for Forward primer is 64.2, and that of reverse primer is 65.5. Can some one suggest how to get the best annealing temperature? Thanks...

01 March 2021 360 7 View

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 3 View

Anybody with an idea on how do design specific primers for detecting tomato resistance genes tm-1, TM-2 and Tm2^2?

I am trying to identify these 3 genes among some tomato cultivar collections and after aligning some sequences from NCBI, I couldn't find unique sequences to target for specific primers. There...

28 February 2021 606 3 View

What is the ideal efficiency for a qPCR?

hello everyone, I need to do standard curves for my qPCR, what is the ideal efficiency range? I tried a primer (Mglu2 receptor) that gave an efficiency of 90.2%. Is it accepted?

28 February 2021 1,254 3 View

Genome-wide gene identification protocol ?

Is there any book chapter/book, webpage or research article available to understand genome-wide gene identification?

28 February 2021 8,095 1 View