Bacterial Transformation in Agrobacterium Rhizogenes ?

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How to we see transcription activity of the bhlh79 family is at the N-terminal or C-terminal?

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Hi there, someone has the SeinFit software for windows because I cannot download it?

DOS version.

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In a multi-scale calculation using ORCA, are charges of the QM system updated during each iteration of optimization?

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How can machine learning be employed in the field of natural sciences?

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Cell Culture Debris? or Contamination?

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How to confirm the site-directed mutagenesis result without performing NGS?

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E.coli contamination in human RNA seq data ?

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Why is my ASO degraded quickly on agarose gel?

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Where should the ARS sequence be introduced on a plasmid?

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Why after performing site directed mutagenesis ,I don't see any colony after transformation?

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Why my colony PCR results of my recombinant bacterial not showing any results?

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How is the bacterial genome's high protein count verified as genuine despite 800+ contigs and good metrics (98.55%completeness, 0.68% contamination)?

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For the moringa oleifera extract using ethanol as the solvent,what is the alternative method to concentrate the extract other than rotary evaporation?

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Transfection in HEK293T cells?

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Mohammed Al-Shakarchi

Try to measure the absorbance using a nanodrop device as a first step to know the concentration and purity of DNA. In the case of a good concentration, carry out the migration process, and if not, increase the amount of additive from bacterial cultures

Ashok Kumar

Recheck your whole process.

Nikolay Klyashtornyy

Some troubleshooting steps you can take:

Check the quality of your DNA extraction reagents: Ensure that the reagents you are using for plasmid isolation, such as isopropanol, DEPC water, RNase A, absolute alcohol, and ammonium acetate, are of high quality and have not been contaminated. It's important to use RNase-free reagents to avoid degradation of your DNA.

Verify the efficiency of plasmid isolation: Double-check your plasmid isolation protocol to ensure that all steps, including lysis, precipitation, and washing, are performed correctly. Pay special attention to the lysis step, as inefficient lysis can result in poor DNA yield.

Assess the integrity of your DNA: It's possible that your DNA may have been degraded during the plasmid isolation process. To check for DNA integrity, you can run a control sample of known DNA on the gel alongside your experimental samples. If the control DNA shows proper bands, it indicates that the problem lies with your plasmid isolation procedure.

Optimize the plasmid isolation protocol: If you consistently observe low or no DNA yield, you may need to optimize your plasmid isolation protocol. Consider modifying the lysis conditions, such as extending the lysis time or using alternative lysis buffers. Additionally, adjusting the centrifugation speeds and times during the precipitation steps can improve DNA recovery.

Verify the sensitivity of your gel electrophoresis setup: Ensure that your gel electrophoresis apparatus, power supply, and staining/visualization methods are functioning properly. Confirm that the gel and running buffer are prepared correctly and that the appropriate voltage and running time are used.

Repeat the plasmid isolation process: If none of the above steps yield satisfactory results, you may need to repeat the plasmid isolation process from the beginning. Pay careful attention to each step, ensure proper handling of the samples, and use freshly prepared reagents.

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