Re: PCR product storage

Once I have a purified PCR product, what temperature is best for storage prior to sequencing? How long can it be stored at that temp?

It depends what the PCR products are stored in. If it is in water it is best to freeze the sample unless you are going to use them within a few hours.

I prefer to use 10/0.1 TE [10mM Tris, 0.1 mM EDTA, pH 8.5] to store my PCR products. The tris helps stop the solution from becoming acidic (cause by CO2 being absorbed from the air) and the small amount of EDTA doesn't interfere with the sequencing reaction. If you use this then you can just store at 4.deg.C. I have found that stored in 10/0.1 TE the PCR products will last for years in the fridge.

Daniel Tillett
Nucleics Support

Pls let me know the condition to store the RAPD PCr product in the refrizarator for how many days
thanks

may i know why PCR products cannot be stored for an extended time if they are not properly sealed and preserved?

Lots of reasons, but the basic issue is they are not sterile. Bacteria will eventually grow and produce nucleases that chew up all the PCR products.

Daniel Tillett
Nucleics Support

Is it better to keep PCR product for long or we should keep them in peg purified form. How long can we keep our PCR products.

How long we can keep our PCR products(if they are in water) in the fridge before purificatin procedue and how long before cloning? Don't freezing and thawing influence our tests? Do you recommend us to freeze all of our samples in water after PCR,if we don't have time to continue the procedure immediately?

The problem with storing PCR products in water is if you have any nucleases around. It is better to store them in 10/0.1 TE. This way the pH won't drop from absorbance of CO2 and the EDTA will keep the nucleases from chewing up the DNA.

Daniel Tillett
Nucleics Support

Jinous Wrote:
-------------------------------------------------------
> How long we can keep our PCR products(if they are
> in water) in the fridge before purificatin
> procedue and how long before cloning? Don't
> freezing and thawing influence our tests? Do you
> recommend us to freeze all of our samples in water
> after PCR,if we don't have time to continue the
> procedure immediately?
My refrigerator (-20 c) was switched off for overnight and my PCR products were there. Is it o.k ?

After having mixed together the master mix along with primers, templates, et cetera, how long (and how) can I store the tubes BEFORE running PCR?

Hi, I have the same question.

Hey,
after PCR how can I storage product?

Hi Kate

In the fridge short term and in the freezer long term. It is not a good idea to store PCR products if you plan to TA clone - storage seems to remove the 3' A overhang.

Daniel Tillett
Nucleics Support

Hi, Why the 3' overhangs tends to be removed after long time storage in the freezer? Someone have any paper that say that or is just popular culture? Thanks

This is lab forklore, but I can say I have tested it myself and it true.

Daniel Tillett
Nucleics Support

hi
my issue is also related to PCR storage product, the problem is that I am doing my PCRs and finnlizing before purification and sequencing, and have to send the sample after PCR (without any purification) to the lab for purification as well sequencing (as v hav't both of these right now in our lab), thus please tell how long I could store my PCRs product and at what temperature??and at what way best to send the samples safely to outside country...



anxiously waiting for kind response