Hello i am a first timer in extracting bacterial dna from soil, and the two attempts i did was all failed during the gel electrophoresis?

here could be several reasons why you did not get the expected results. Here are some possible causes:

Low yield of DNA extraction: Soil contains a diverse range of microorganisms, including bacteria, fungi, and archaea. The DNA extraction protocol that you used may not have effectively extracted DNA from all the bacterial species present in the soil. It's possible that the yield of DNA was too low to detect on the gel.

Inhibition of PCR amplification: Even if you were able to extract DNA from the soil, the presence of inhibitors, such as humic acids and other organic compounds, may have inhibited PCR amplification. Inhibition can result in low amplification efficiency or even no amplification at all.

Poor quality DNA: The DNA that you extracted may have been of poor quality, which can affect the efficiency of PCR amplification. Factors that can affect DNA quality include incomplete cell lysis, degradation of DNA during extraction or storage, and the presence of contaminants.

To troubleshoot your experiments, you can try the following:

Optimize your DNA extraction protocol: Try using a different DNA extraction kit or modify the protocol to improve the yield of DNA extraction.

Purify your DNA: After DNA extraction, purify your DNA using a commercial DNA purification kit or by using phenol-chloroform extraction and ethanol precipitation to remove any inhibitors or contaminants.

Optimize PCR conditions: Optimize your PCR conditions, such as annealing temperature, extension time, and primer concentrations to improve PCR amplification efficiency.

Use positive and negative controls: Include positive and negative controls in your experiment to check for contamination, DNA quality, and amplification efficiency.

Verify the identity of your bacteria: Verify the identity of your bacteria using 16S rRNA sequencing to confirm that you are amplifying the correct target.

I hope these tips help you troubleshoot your experiment and achieve successful results in your future attempts. Good luck!

These video playlists might be helpful to you:

https://www.youtube.com/playlist?list=PLzBiAPKi8vOPbE7Db9mKwloe34E8rXXEU

https://www.youtube.com/playlist?list=PLzBiAPKi8vONSqjcweFWHMRy13y3484bE

https://www.youtube.com/playlist?list=PLzBiAPKi8vOMheKwb0mu1dcZRaeGUFSlw

You may also try these kits:

PowerSoil DNA Isolation Kit: This kit is designed to extract DNA from a wide range of environmental samples, including soil. It uses bead-beating technology to efficiently lyse cells and release DNA.

MoBio PowerMax Soil DNA Isolation Kit: This kit also uses bead-beating technology to extract DNA from soil samples. It includes a step for removing humic acids, which can interfere with downstream applications.

ZymoBIOMICS DNA Miniprep Kit: This kit is designed to extract DNA from a variety of environmental samples, including soil. It uses a combination of bead-beating and chemical lysis to efficiently lyse cells and release DNA.

Omega Biotek E.Z.N.A. Soil DNA Kit: This kit uses a combination of mechanical and chemical lysis to extract DNA from soil samples. It includes a step for removing humic acids, which can interfere with downstream applications.

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