RFLP Problem (View forum version)



sidra_younis

Posted 01 January 2013 - 04:18 AM

I am facing problem with restriction digestion of my pcr amplified products. i get good DNA. Good pcr product but in RFLP some of sampels digest properly and some remain undigested or partially digested. I am attaching pcr and rflp gel. Any one please help me.  rflp.docx   26.13KB   433  rflp.docx   26.13KB   433  rflp.docx   26.13KB   433

phenol chloroform method for dna isolation. pcr for il 1 gene....and rflp using hin 111 enzyme. many of my colleagues have used same protocols for same primers and they got good results. Even with some (hundred) of my own samples gave good digestion with same primers and enzymes...but now something is hindering digestion....

sidra_younis

Posted 01 January 2013 - 04:27 AM

Dear all...I am facing problem with restriction digestion of my pcr amplified samples. i get good dna pellet..good pcr product...but during restriction digestion some of samples are properly digested...either remain undigested or incompletely digested...i am attaching rflp gel pic...Please someone guide me


phage434

Posted 01 January 2013 - 09:12 AM

No possible way we can help without far more information. We are not mind readers. Protocol. Target. Primers. Enzymes.