| | |||||||
| Register | Search | Today's Posts | Mark Forums Read |
| Real-Time PCR and Quantitative PCR Forum Real-Time PCR and Quantitative PCR Forum |
| | LinkBack | Thread Tools | Display Modes |
|
#1
| |||||||||||
| |||||||||||
| I have many samples containing a bacterial genomic DNA. I want to measure the quantity of 7 unique sequences within the genomic DNA and compare them to each other within the same sample. I am thinking that absolute quantification using a standard curve is the best method to use. Due to the number of primer sets (7), setting up a standard curve on each plate would not be possible using all 7 primer sets. Is it advisable to set up a standard curve for 1 sequence (A1) and then measure the amount of A1 in 26 samples. This would be repeated for sequences A2-A7. Is one able to then compare the amount of A1-A7 sequence in the same sample (S1) across the 7 plates? I think this might be ok since you are no longer comparing Ct's, but you are comparing copy number based off of the standard curves. Would a relative standard curve or efficiency-corrected delta-Ct be any better? I don't think the relative standard curve would be needed since I am not comparing values across samples, just the number of unique sequence tags within each sample. Thanks for any advice. I've only used qPCR as a measure of gene expression in the past, so am not as sure how to just quantify tags within the same sample of DNA. |
| Tags |
| dna , method , quantification |
| Thread Tools | |
| Display Modes | |
|
|
| | ||||
| Thread | Thread Starter | Forum | Replies | Last Post |
| qPCR NEWS April 2009 - focus on microRNA normalisation | Editor www.Gene-Quantification.info | Protocols and Methods Forum | 0 | 04-29-2009 02:18 PM |
| qPCR NEWSLETTER - July 2007 | Editor www.Gene-Quantification.info | Protocols and Methods Forum | 0 | 07-27-2007 11:04 AM |
| qPCR NEWSLETTER - October 2006 | Editor www.Gene-Quantification.info | Protocols and Methods Forum | 0 | 10-26-2006 12:25 PM |
| Cylindrical wave, wave equation, and mistakes | h_v_ansari@yahoo.com | Physics Forum | 2 | 10-26-2006 01:47 AM |
| Scientific method (was Re: Fusion In Utah (again)) | Bill Vajk | Chemistry Forum | 12 | 09-29-2003 08:30 PM |