3 replies to this topic

[Bassaml7]

Hi ,

I just have a couple of questions regarding storage of some PCR components.

1) Is it better to dilute and store dNTPs in just water or PCR buffer 1X ?

I would prefer PCR buffer because water can be slightly acidic and also carbondioxide from air may dissolve in dNTP solutions causing PH to decrease further . Low PH causes dNTP hydrolysis as far as I know, so I'd think storing dNTPs in a buffered solution would be the best Practise . But yet I found too many using just water . So I'd like to share thoughts about that . What do you think is best ?

2) How many reactions should a single aliquot of dNTPs \ Primers be sufficient for ? How many freeze\thaw cycles do you think dNTPs \ Primers can withstand before problems can occur ?

3) I heard that Taq could be stable at 2-8 C for about 2 weeks ? is that true ?


Thanks for your time.

Edited by Bassaml7, 05 September 2009 - 07:40 AM.

[leelee]

I dilute my dNTPs in nuclease free water, never had a problems with them. I make up around 100ul, and this usually lasts me several months, and plenty of freeze-thaws with no probs.

[Bassaml7]

leelee, on Sep 5 2009, 07:33 PM, said:

I dilute my dNTPs in nuclease free water, never had a problems with them. I make up around 100ul, and this usually lasts me several months, and plenty of freeze-thaws with no probs.

thank you very much for your reply . That helps relieve my stress a bit.

[jiajia1987]

Bassaml7, on Sep 5 2009, 10:20 PM, said:

Hi ,

I just have a couple of questions regarding storage of some PCR components.

1) Is it better to dilute and store dNTPs in just water or PCR buffer 1X ?

I would prefer PCR buffer because water can be slightly acidic and also carbondioxide from air may dissolve in dNTP solutions causing PH to decrease further . Low PH causes dNTP hydrolysis as far as I know, so I'd think storing dNTPs in a buffered solution would be the best Practise . But yet I found too many using just water . So I'd like to share thoughts about that . What do you think is best ?

2) How many reactions should a single aliquot of dNTPs \ Primers be sufficient for ? How many freeze\thaw cycles do you think dNTPs \ Primers can withstand before problems can occur ?

3) I heard that Taq could be stable at 2-8 C for about 2 weeks ? is that true ?


Thanks for your time.

I dilute and store my dNTPs in just water. Keep it up to 200uL at times and have no problems with it. The same goes for primers. As for Taq, I always keep it at -20degcel, never at 2 to 8degcel.