{"id":9694,"date":"2019-07-30T15:07:08","date_gmt":"2019-07-30T21:07:08","guid":{"rendered":"https:\/\/www.coleparmer.com\/blog\/?p=9694"},"modified":"2019-08-06T14:19:34","modified_gmt":"2019-08-06T20:19:34","slug":"is-a-4c-holding-temperature-for-a-pcr-reaction-necessary","status":"publish","type":"post","link":"https:\/\/www.coleparmer.com\/blog\/is-a-4c-holding-temperature-for-a-pcr-reaction-necessary\/","title":{"rendered":"Is a 4\u00b0C Holding Temperature for a PCR Reaction Necessary?"},"content":{"rendered":"

By Michael Steinert, Product Manager, Biosciences, Cole-Parmer<\/p>\n

Determining what\u2019s best for your experiment vs. your thermal cycler<\/h2>\n

As a research scientist in my past life, I have been in the position as many others, where I simply left a completed plate of PCR reaction in a thermal cycler overnight\u2014not at a holding temperature of 4\u00b0C or 10\u00b0C, but at room temperature. My immediate thought was that the reaction was ruined and that successful genotyping was impossible. However, to my surprise, the gel electrophoresis run with this \u201cruined\u201d reaction provided the necessary separation and band concentration I needed to genotype my mice.<\/p>\n

As it turns out, DNA is extremely stable even at room temperature for a short duration, and some studies suggest it can remain stable for days. This is why many in the scientific community use a holding temperature of 10\u00b0C instead of 4\u00b0C and why many newer thermal cyclers do not even allow for 4\u00b0C hold. There a few reasons for this, and the main argument is the stability of a completed PCR reaction allows for the high integrity of the reaction at 10\u00b0C while also putting significantly less strain on your thermal block.<\/p>\n

Having the block at a low temperature such as 4\u00b0C can actually cause more condensation than a higher holding temperature. This condensation can actually cause damage to the block overtime, significantly reducing the lifespan of the machine. Not to mention, by simply setting your temperature that low, you are already requiring more work from your equipment. A 10\u00b0C hold will cause less wear and tear on your machine and lead to a piece of PCR equipment that runs without issues for years to come. However, for long term storage, it is best to store your completed reaction at -20\u00b0C to preserve the sample.<\/p>\n

Of course, if you have been using a 4\u00b0C hold for much of your scientific life, I can see why you would be hesitant to give the 10\u00b0C hold a try. If you have enough sample, and time to compare, I would suggest seeing if the 10\u00b0C vs. 4\u00b0C hold shows any difference in yield or band visibility. If it doesn\u2019t, you may have saved years of life for your thermal cycler, and saved thousands of dollars in the long run.<\/p>\n

Learn more about Simplified Optimization with PCRmax Alpha Cyclers<\/a><\/p>\n

Browse the selection of Alpha Thermal Cyclers<\/a> available from Cole-Parmer<\/p>\n

References:<\/h3>\n

https:\/\/www.colorado.edu\/ecenter\/sites\/default\/files\/attached-files\/seracare_stability_of_genomic_dna_at_various_storage_conditions_isber2009.pdf<\/a><\/p>\n

Jennifer Joiner, Molecular Staging Inc. New Technology Increases the Availability of High Quality DNA for Genetic Testing. CSR News, 2002 2.John G. Baust, Strategies for the Storage of DNA Biopreservation and Biobanking 6:251\u2013252 (2008) 3.Lasken RS, Egholm M. Whole genome amplification: abundant supplies of DNA from precious samples or clinical specimens. Trends Biotechnol. 2003;21:531\u201353<\/p>\n

John G. Baust, Strategies for the Storage of DNA Biopreservation and Biobanking 6:251\u2013252 (2008)<\/p>\n

Lasken RS, Egholm M. Whole genome amplification: abundant supplies of DNA from precious samples or clinical specimens. Trends Biotechnol. 2003;21:531\u2013535.<\/p>\n","protected":false},"excerpt":{"rendered":"

By Michael Steinert, Product Manager, Biosciences, Cole-Parmer Determining what\u2019s best for your experiment vs. your thermal cycler As a research scientist in my past life, I have been in the position as many others, where…<\/p>\n","protected":false},"author":20,"featured_media":9702,"comment_status":"open","ping_status":"open","sticky":false,"template":"","format":"standard","meta":{"footnotes":""},"categories":[1964],"tags":[],"class_list":["post-9694","post","type-post","status-publish","format-standard","has-post-thumbnail","hentry","category-pcr"],"yoast_head":"\nIs a 4\u00b0C Holding Temperature for a PCR Reaction Necessary? | Cole-Parmer blog<\/title>\n<meta name=\"description\" content=\"If you've been using a 4\u00b0C hold for a PCR reaction, give a 10\u00b0C hold a try. You may save years of life for your thermal cycler and thousands of dollars. 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