Search found 45 matches

by bravebeaker
Wed Oct 24, 2012 2:05 am
Forum: Site Suggestions
Topic: Tapatalk App iOS/Android
Replies: 0
Views: 8375

Tapatalk App iOS/Android

Dear forum admins, Hi.. Nowadays its difficult for anybody to go anywhere without keeping his/her cell phone close and recently tablets are also been carried almost all the time. I am sure you are aware of an application on iOS and on Android systems called "Tapatalk" Its a very powerful a...
by bravebeaker
Fri Oct 19, 2012 6:26 am
Forum: Molecular Biology
Topic: Real time PCR
Replies: 4
Views: 3776

make sure to vortex your DNA samples before you mix them for the qPCR in order to generate the std curve. also make sure you measured the DNA accurately to make the std curve calculation, for example I found out that checking by nanodrop doesnt give me good results as good as Qubit (different measur...
by bravebeaker
Wed Oct 17, 2012 3:56 am
Forum: Molecular Biology
Topic: Primers Tm
Replies: 7
Views: 5575

Thank you for your advice I will give the current primers a try and will start editing them. I dont want my sensei to yell at me! :P Thanks alot!
by bravebeaker
Mon Oct 15, 2012 10:46 am
Forum: Molecular Biology
Topic: Primers Tm
Replies: 7
Views: 5575

Today I used gradient PCR and I set the annealing temp to start from 55 and increase 15 degrees gradually. the range was as follows: 55.1 55.5 56.4 57.6 59.2 61.1 63.8 65.7 67.4 68.8 69.9 70.3 I also tried 1:1 ratio and 2:1 (Low Tm primer conc : High Tm primer conc) Both genes were amplified startin...
by bravebeaker
Fri Oct 12, 2012 9:47 am
Forum: Molecular Biology
Topic: Primers Tm
Replies: 7
Views: 5575

can I ask a quick question? I checked the PCR machine I am using now and I can modify the run by adding an increment of .5 degree to each cycle starting from 59 C. but how can I assign a range as you recommended from 59 to 68? is it by using the Rotor Q? because I couldnt find the right option in th...
by bravebeaker
Fri Oct 12, 2012 5:55 am
Forum: Molecular Biology
Topic: Primers Tm
Replies: 7
Views: 5575

Thanks for the reply will do and let you know what happens.
by bravebeaker
Fri Oct 12, 2012 12:44 am
Forum: Molecular Biology
Topic: what is gain in fluorescence detector
Replies: 6
Views: 10144

Re: what is gain in fluorescence detector

care to elaborate how you solved the problem? I used to use Roche light cycler 480 and now I am using RotorQgene and for the RotorQ it optimizes the gain before the run and sometimes during the run. I also found out that the intensity of the fluorescnce depends on how many tubes are in the rotor. I ...
by bravebeaker
Thu Oct 11, 2012 7:18 am
Forum: Molecular Biology
Topic: Primers Tm
Replies: 7
Views: 5575

Primers Tm

Hello everyone, I am having some difficulty determining the optimal Tm for my primers for PCR. I ordered two primers for different genes. gene A primers Tm 59 and 71 gene B primers Tm 64 and 71 I am trying to run them both at the same time since they amplify almost the same size of a product which i...
by bravebeaker
Fri Dec 16, 2011 9:02 am
Forum: Molecular Biology
Topic: Protoplast Isolation - first step - enzyme digestion
Replies: 0
Views: 1881

Protoplast Isolation - first step - enzyme digestion

Hello all, I have been reading lots of papers about protoplast isolation. I found that most of the papers but not all say that enzyme digestion is in the dark. Why should it be in the dark? i have been trying to get an explanation but cant find any. In the mean time I am trying to enzyme digestion w...
by bravebeaker
Tue Dec 06, 2011 8:46 am
Forum: Molecular Biology
Topic: DO NOT UNDERSTAND PRIMER DESIGN
Replies: 4
Views: 5493

I would really like to know too! honestly there are so many ways and methods on web I found. The thing is maybe I dont have the courage to apply it without returning to an experianced person!
Will follow up the thread...
by bravebeaker
Tue Dec 06, 2011 8:43 am
Forum: Molecular Biology
Topic: DNA purity
Replies: 1
Views: 1668

to the best of my knowledge, pure DNA preparation have expected ABS260/ABS280 ratio between 1.8 and 2.0.
by bravebeaker
Thu Oct 20, 2011 3:37 am
Forum: Molecular Biology
Topic: general QPCR problem
Replies: 9
Views: 11347

Re: Re:

thats fine for cDNA. Do you isolate total RNA or message alone, and do you perform any control primers to test for genomic actin? I isolate total RNA and I dont use control primers to test for genomic actin but during my reverse trascriptase step I prepare -ve controls to check for contamination, a...