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PCR Troubleshooting - No Bands or Product After PCR

Troubleshooting PCR No Product No Bands

This is a PCR troubleshooting page for troubleshooting the PCR primer dimers band/product problem and its solution.

No PCR Products or Bands Problem

No PCR Products are usually encountered when you run your PCR reaction on an agarose gel and you visualize no PCR bands after staining.

No PCR Products or Bands Solutions - Troubleshooting Guides

  • USE Freshly ISOLATED DNA Template
  • Use Fresh Reagents and Polymerase
  • Treat for DNAses
  • Use Filtered UV treated / Autoclaved Water
  • Your primers may be annealing to each other. Re-design primers and order a new batch.
  • Make sure you use primer design software and check for self-annealing and that the primers do not share a large percentage of complementary sequence.
  • Check primers carefully for homo-dimer and hetero-dimer formation with OligoAnalyzer
  • Try adding DMSO up to 5%.
  • Try using HotStart PCR instead of regular Taq polymerase PCR.
  • Conduct PCR with and without formamide.
  • Titrate Mg2+ (MgCl - 1.5, 2.0, 2.5 and 3.0 mM) concentration.
  • Increase the DNA template amount (concentration).
  • Increase the annealing temperature (try optimizing using a gradient PCR machine to find optimal temperature for annealing).
  • Use a combination of some/all of the above.

Can't Find the Solution to your PCR Problem? Make sure you visit the PCR Forum for PCR troubleshooting and solutions to common PCR problems. You can also post questions to other researchers by simply registering first and then clicking on Post New Thread button in the forum, or simply clicking the question mark below!

PCR Forum Topics

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Cresol Red/Sucrose Hi. Does anyone have experience using cresol red/sucrose as loading dye added to the reaction BEFORE PCR? Does cresol red/sucrose have to have a...
Amplification won't work! I'm trying to amplify a ~200bp segment from a ~4kb plasmid. A postdoc in my lab previously amplified the segment and gave me a stock of gel-extracted...
Any ideas why my PCR amplification won't work? I'm trying to amplify a ~200bp segment from a ~4kb plasmid. A postdoc in my lab previously amplified the segment and gave me a stock of gel-extracted...
Heated lid on RFLP: is it ok? I'm doing three different RFLPs with three different restriction enzymes using a single amplicon, each RFLP in different tubes of course as each...
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