I want to know for example If delta ct value be 6, is it acceptable for you? in what range delta ct value is unacceptable usually? I know it depends on my target and reference gene and I also know reference gene selection in terms of its expression compared with my target expression is so important
Laurence Stuart Dawkins-Hall
Dear Molood
- a delta Ct of 6 is perfectly ok.
- Of couse it will vary depending on the relative expression of your housekeeper and target gene
- What is important is that for triplicate reactions that value doesn't vary by more than +/-0.5 cycles
- Also for both target and Housekeeping genes make sure that your Ct values used to derive your Delta Ct value are between (ideally) 20-30cycles. For house keepers you can go below 20 cycles (providing that value is consistent) but try not to go above 30 cycles for your gene of interest
- Providing your delta Ct value is derived in line with the above criterion a delta Ct value of 6 is perfectly acceptable
2 votes
1 thanks
Ahmed Abdelfattah
Dear Molood
Ct value of 6 is very high but acceptable with no doubt.
if you are not comfortable with it, you can increase the Ct value by diluting the target and reference gene 1:10 or 1:100 using the same experimental procedures.
all the best
0 votes
0 thanks
- Badges
- Science method
More Molood Behbahanipour's questions See All
Similar questions and discussions