Dear all, what can happen if I put NEB-end-repair reaction in 30C (30') instead of 20C?

How to calculate the fraction metabolized of a drug by an enzyme using clearance(ml/min) values of the metabolite formed?

I am trying to find the fraction metabolized of a drug by CYP and UGT enzymes. I am unable to get the values of fraction metabolized (Fm) directly from the literature. Can we calculate the Fm of a...

21 February 2021 5,379 2 View

How long does it take to design and validate transition state analogue inhibitors?

Hi, I'm asking from the point of view of designing a research plan: How long (approximately) can it take to determine the transition state structure of an enzyme substrate using kinetic isotope...

14 February 2021 2,610 1 View

Kinetics of Enzyme production in Recombinant E Coli?

Hello, I am trying to find kinetic information (such as rate of enzyme production etc.) on the expression of taq polymerase in E-Coli after inserting a recombinant plasmid. Any help would be...

12 February 2021 1,386 3 View

No title

Hello! I want to determine the activity of my in house Reverse Transcriptase enzyme. For this, I have used Thermo's Enzcheck Reverse Transcriptase assay kit. I used commercial Thermo RT enzyme to...

08 February 2021 7,285 1 View

How define one baker unit(BU) is the amount of hydrogen peroxide catalase ?

07 February 2021 2,006 1 View

Does DNA A-tailing survive after agarose gel electrophoresis and purification?

Hi everyone, I need to clone in pGEM-T vector the result of amplifying a metagenomic library by PCR with a high fidelity polymerase but I only want to clone the fragments of an specific size...

04 February 2021 6,051 8 View

What are some common oxidants used to oxidize iron sulfur clusters in proteins for techniques like EPR?

I have used potassium ferricyanide (which is likely the most common) extensively in the past, but I am curious to know if there are other oxidants that work well. I need to generate the oxidized...

29 January 2021 9,803 4 View

Papain Enzyme Assay...?

Hi all, I would like to know how can I estimate the activity of the enzyme, papain, which is used in reactions as crude latex extract of papaya. Please share some procedures if anybody having...

26 January 2021 569 4 View

Where can I find plants pyr-GC-MS data?

25 January 2021 5,403 2 View

Recombinant restriction enzyme: how strong should the corresponding methyltransferase expression level be?

Hello. We are planning expression of a restriction enzyme in E. coli, and wondering how much of the corresponding methyltransferase is needed to be expressed in order to protect the host genome...

23 January 2021 2,096 4 View

N. Scott Reading

Check the literature from NEB, write a note to NEB and determine if there is a temperature at which this enzyme will inactivate or have some other form of activity. The most likely reaction is that the enzyme activity was lower than optimum but it was still functional.

However, since you did the experiment only you are in the best position to evaluate how the reaction performed by following through with the remainder of what you had planned. (Or you may just scrap the current effort and repeat the process correctly.)

Good luck, let us know how it worked out.

Micha Ben-Zimra

Thanks!

Dear Peter,

Indeed, according to Q-bit concentration of DNA at the end of the process it looks like it worked. Is it the first enzyme or the second addition of 0.5 ul?

Micha