What happends if you store your ligation reaction at room temp for too long? (Plasmid+vector+t7 ligase and buffer)?

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Is HOECHTS/PI staining (and then quantify) a good assay to determine the amount of dead cells of a whole population?

I want to treat fibroblasts with 24h MG132 and then look a the amount of cell death. However, I want to correct for the amount of cells in total, because they also prolfirate in 24hour. My...

10 November 2018 9,018 3 View

AMAXA Lonza nucleofection lower volume of buffer?

Does anyone tried to do nucleofection with AMAXA by Lonza with lower than recommended amount of buffer in the cuvettes (100 ul)?

07 August 2024 4,616 0 View

Why I can't see any band in SDS-PAGE?

Currently, when I run SDS-PAGE, I don't see any bands at all, even though I used the same material just a day ago and it worked fine.... In our lab, we dilute the 10X running buffer to 1X and...

06 August 2024 5,373 2 View

Why might the impedance values for DI water and 0.1X PBS buffer solution exhibit a decreasing and increasing trend, respectively over time (HP 4194A)?

Hello everyone, I'm encountering an issue with my electrochemical impedance spectroscopy (EIS) measurements and would appreciate some insights. Experimental Setup: Electrodes: Gold interdigitated...

05 August 2024 3,783 2 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

What is the best blank for nanodrop if I want to read a recombinant protein concentration?

Is it the "elution buffer" or the "dialysis buffer"? Note: I'll be using NanoDrop OneC

01 August 2024 967 3 View

EDTA buffer solution?

Hello everyone! How can i prepare EDTA buffer solution (pH 8.2) to determine GSH levels? Thank you in advance.

29 July 2024 4,374 1 View

Whether i can preserve leaves samples in 2.5% glutaraldehyde in 0.05 M phosphate buffer, pH 7 for coming several weeks for further LM study ?

I want to preserve the sample leaves for the LM study of the stomata and conidia analysis. Whether 2.5% glutaraldehyde in 0.05 M phosphate buffer with pH 7 can be applied for preservation.

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Can a shoot-through event of a tri-state digital buffer cause momentary Hi-Z state?

// interested in the difference between floating events and short circuits.

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How can EDTA be removed from already extracted DNA samples?

Our lab has extracted bacterial DNA samples using Qiagen DNeasy kit and eluted the samples in AE buffer. However, this buffer contains 0.5mM EDTA not suitable for downstream application. Any ideas...

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How much 1X TAE buffer should I use in preparing 1.2% agarose gel?

Commonly, we prepare 1.0% of agarose gel with 0.4g agarose powder and 40ml 1X TAE buffer. if I would like to prepare a 1.2%% of agarose gel, is it I need to add 0.48g agarose powder and 120ml 1X...

12 July 2024 2,058 5 View

Dominique Liger

Hi there,

Apart from losing ligase activity I don't think much happen to the ligation product if the ligation mix is free of any DNA degrading activity.

Steingrimur Stefansson

..and it has no bugs growing in it.

Shamsher S. Kanwar

Ligase activity shall be lost and the plasmid & DNA may gedrade by nucleases activity of contaminating microbes!