Javascript Disabled Detected

You currently have javascript disabled. Several functions may not work. Please re-enable javascript to access full functionality.

kyakhoob

Member Since 27 Sep 2012
Offline Last Active Oct 10 2012 11:31 PM

Find content

Community Stats

Group Active Members
Active Posts 6
Profile Views 183
Member Title member
Age Age Unknown
Birthday Birthday Unknown
Gender
Not Telling

About me

My research interests
molecular biology

Contact Information

0 Neutral

Friends

kyakhoob hasn't added any friends yet.

Latest Visitors

hobglobin
09 Oct 2012 - 07:08

Posts I've Made

In Topic: nonspecific band at 50bp in PCR

10 October 2012 - 11:31 PM

hobglobin, on 10 October 2012 - 08:32 AM, said:

well I actually do it when they were quite long in the fridge, because when using them frequently, the condensing water is no problem...anyway with frozen and then thawed solutions mixing is more important to avoid heterogeneous concentrations within the tube...especially with MgCl₂ and buffer....and after this, the centrifuging is not necessary of course.

Ok..I am going to try it again..... fingers crossed ...

In Topic: nonspecific band at 50bp in PCR

09 October 2012 - 10:25 PM

hobglobin, on 09 October 2012 - 07:52 AM, said:

and the DNA was ok? and I didn't get what centrifuging primer working solution has to do with thawing? I store my working solutions in the fridge and centrifuge them shortly before using, to get down the condensed water and mix it. The same with other solutions from the fridge where this problem may occur.

'hobglobin
yes my DNA is ok. As you said that that "your primer working solutions are frozen as several diluted aliquots from the stock. as, you store your working solutions in the fridge and centrifuge them shortly before using, to get down the condensed water and mix it. " my question was that should I centrifuge all solution (Buffer, MgCl2, dNTP) including my primer working solution????
I used to centrifuge only the MgCl2 before using it. and i use freshly prepared dNTP each time. Is it important to centrifuge the primer working solution before using it???
please suggest me.

In Topic: nonspecific band at 50bp in PCR

09 October 2012 - 03:46 AM

hobglobin, on 28 September 2012 - 05:41 AM, said:

kyakhoob, on 28 September 2012 - 04:03 AM, said:

habglobin

thank you for your suggestion actually i have tried everything. i have added every reagent properly..i tested the DNA and found OK..primers are few month old. my guide told me that the primers might get contaminated. is there any way to know whether the primers are okay or not. please help.

how did you check the DNA then? and primers, I'd also replace, i.e. throw away the working solutions and thawing new ones (my primer working solutions are frozen as several diluted aliquots from the stock).
also checking the thermal cycler is an idea, i.e. asking if colleagues have no problems with their PCRs and if your program is still untouched...

Habglobin
sorry for late reply.
Thank you for your suggestion. I checked the DNA by running genomic DNA in 0.8% gel. I checked the thermocycler and it is OK. Can I centrifuge the the working primer solution before preparing themaster mix to avoid thawing of primers....???? please. help

In Topic: nonspecific band at 50bp in PCR

09 October 2012 - 03:45 AM

Habglobin
sorry for late reply.
Thank you for your suggestion. I checked the DNA by running genomic DNA in 0.8% gel. I checked the thermocycler and it is OK. Can I centrifuge the the working primer solution before preparing the master mix to avoid thawing of primers....???? please. help

In Topic: nonspecific band at 50bp in PCR

28 September 2012 - 04:03 AM

Google Sign in options