To establish a method of semiquantitative polymerase chain reaction for detecting CdC 6 gene / 中华检验医学杂志

ABSTRACT

Objective To establish a method of semiquantitative polymerase chain reaction for detecting cdc6 gene in EJ cells and bladder cancer tissue.Methods We collected the samples of urine EJ bladder cancer cells and extracted the total RNA, and then did RT-nest-PCR to check the expressive level of cdc6 gene. Result We established stable semiquantitative PCR by putting primer of beta-actin served as internal reference gene and primer of target gene in the same test tube and optimizing experiment parameters for PCR, such as the concentration of magnesium and cycle times etc. The intro and inter group CV were 8.01 and 14.53 respectively for cdc6. The detecting limit was 5?10 -2 ng.Conclusion It is usable to test the expressive level of cdc6 gene in bladder cancer.