Hi,
In our lab, we are working with qPCR, for mutation detection. Now the problem is that all of our Cp-values, for our test DNA, are too high. The values are over Cp: 32 and more, for our test DNA. We don't know what the problem is, since we used purified DNA (so it can't be that its because of contamination, I think).
Because of this, we don't dear to start, with mutationdection of our samples.
Do somebody now, what kind of problem can cause this effect? This would certainly help,
thanks,
Susan
0
2 replies to this topic
#2
Flyingbike
-
- Active Members
-
- 11 posts
member
0
Neutral
Neutral
Posted 12 October 2009 - 05:03 AM
susanna, on Oct 8 2009, 05:06 PM, said:
Hi,
In our lab, we are working with qPCR, for mutation detection. Now the problem is that all of our Cp-values, for our test DNA, are too high. The values are over Cp: 32 and more, for our test DNA. We don't know what the problem is, since we used purified DNA (so it can't be that its because of contamination, I think).
Because of this, we don't dear to start, with mutationdection of our samples.
Do somebody now, what kind of problem can cause this effect? This would certainly help,
thanks,
Susan
In our lab, we are working with qPCR, for mutation detection. Now the problem is that all of our Cp-values, for our test DNA, are too high. The values are over Cp: 32 and more, for our test DNA. We don't know what the problem is, since we used purified DNA (so it can't be that its because of contamination, I think).
Because of this, we don't dear to start, with mutationdection of our samples.
Do somebody now, what kind of problem can cause this effect? This would certainly help,
thanks,
Susan
I think this means a low efficiency PCR. Are your parameters (temperature, elongation time, etc) adjusted according to your amplification target ?
#3
susanna
-
- Active Members
-
- 79 posts
Enthusiast
0
Neutral
Neutral
Posted 15 October 2009 - 04:06 AM
Yes, but i wondered why i have such a low pcr-efficiency.
Were using a kit, so everythings already optimalized.
greetz
susan
I think this means a low efficiency PCR. Are your parameters (temperature, elongation time, etc) adjusted according to your amplification target ?
Were using a kit, so everythings already optimalized.
greetz
susan
Flyingbike, on Oct 12 2009, 06:03 AM, said:
susanna, on Oct 8 2009, 05:06 PM, said:
Hi,
In our lab, we are working with qPCR, for mutation detection. Now the problem is that all of our Cp-values, for our test DNA, are too high. The values are over Cp: 32 and more, for our test DNA. We don't know what the problem is, since we used purified DNA (so it can't be that its because of contamination, I think).
Because of this, we don't dear to start, with mutationdection of our samples.
Do somebody now, what kind of problem can cause this effect? This would certainly help,
thanks,
Susan
In our lab, we are working with qPCR, for mutation detection. Now the problem is that all of our Cp-values, for our test DNA, are too high. The values are over Cp: 32 and more, for our test DNA. We don't know what the problem is, since we used purified DNA (so it can't be that its because of contamination, I think).
Because of this, we don't dear to start, with mutationdection of our samples.
Do somebody now, what kind of problem can cause this effect? This would certainly help,
thanks,
Susan
I think this means a low efficiency PCR. Are your parameters (temperature, elongation time, etc) adjusted according to your amplification target ?
