|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|
is higher Ct for internal control compared to the target gene is acceptable?
I am doing real time PCR using SYBR green chemistry. The primers i m using for endogenous control is elongation factor alpha which i designed form rice sequence. My host plant is a wild relative of rice which have 96% sequence similarity with rice. I always get higher Ct value (nearly 32)for endogenous control gene comapared to the target gene (nearly 23). when i use genomic DNA as a template the amplification for elongation factor is much higher as compared to the amplification obtained using cDNA as a template.what could be the reason for getting such a high Ct value for cDNA. Is my primer for elonagtion factor is not properly binding to the cDNA , because i m getting contracdictory results now with respect to my semiquantitative result. For semiquantitative pcr i used to use the same elongation factor primer and used to get good amplification.i have attached a photo of semiquantitative pcr. the lower band refer to the elongation factor. what could be the possible solution for this problem ?
|acceptable , compared , control , gene , higher , internal , target|
|Thread||Thread Starter||Forum||Replies||Last Post|
|Human Cytome Project - Update 24 Jan. 2005||Peter Van Osta||Cell Biology and Cell Culture||1||08-01-2010 02:18 PM|
|New Saccharomyces Sequences 11/27/04||Mike Cherry||Yeast Forum||0||11-28-2004 11:39 PM|
|New Saccharomyces Sequences 09/08/04||SGD Sequences||Yeast Forum||0||09-13-2004 10:07 PM|
|New Saccharomyces Sequences 08/11/04||SGD Sequences||Yeast Forum||0||08-12-2004 12:26 AM|
|New Saccharomyces Sequences 05/19/04||SGD Sequences||Yeast Forum||0||05-23-2004 04:06 PM|