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-20 freezer fail-- please help!!

-20 freezer fail-- please help!! - PCR - Polymerase Chain Reaction Forum

-20 freezer fail-- please help!! - PCR - Polymerase Chain Reaction Forum. Discuss and ask questions about PCR troubleshooting, PCR protocols and methods, PCR products, and PCR theory.


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Old 10-17-2011, 01:31 AM
Pipette Filler
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Default -20 freezer fail-- please help!!



Hi all,

I went into lab last Friday morning to find that sometime Thursday night (presumably after 6pm or so), our -20C freezer stopped working. Consequently, a bunch of my aliquoted chemicals/PCR supplies were sitting at 4C or higher for up to ~ 12 hours (everything was thawed and there was a huge puddle of water under the freezer). The PCR amplification I tried on Friday failed, but I'm not sure whether it was bound to fail or whether it was my reagents (gonna test with someone else's PCR reagents this week). So I'm trying to decide which aliquots are still good and which I should throw out. I'm fairly sure my plasmid stocks/PCR primers are okay, but everything else is up in the air. Would appreciate your input on whether the following items could survive 12 hours at > 4C:

antibiotic stock solns (ampicillin in water, kanamycin in water, chloramphenicol in ethanol, tetracycline in ethanol)
Pfu and Vent polymerases and their buffers
dNTPs
T4 DNA ligase and T4 ligase buffer
IPTG (1M in water)
actinomycin D stock soln
MTT stock soln
Laemmli sample buffer (with and without DTT)
Dpn I restriction enzyme

I'd really rather not throw out anything more than I have to, but I'd also rather not risk having stuff fail because of bad reagents. Any insight you guys could give would be great!

Thanks!!
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Old 12-10-2011, 05:48 AM
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Default Re: -20 freezer fail-- please help!!

dear sir
dont woried every thing will be alright, this was happened more than once
the reagent will work in my responsibility
mahmoud manama
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