The objective of this study was to examine the effects of presynchronization and recombinant bovine somatotropin (rbST) on conception rates following a timed insemination (TAI) protocol in lactating dairy cows. A further objective included the evaluation of the efficacy of the Early Conception Factor (ECF) test kit. Recombinant bST may offer some benefit when used in conjunction with estrus synchronization and TAI. Presynchronization treatment consisted of two injections of PGF2α given 14 d apart, with the second dose administered 14 d prior to the initiation of a TAI protocol. A total of 216 lactating Holstein cows were presynchronized with PGF2α and then received GnRH (100μg) at 67 ± 7 d post partum (PP), administration of PGF2α (25 mg) 7 d later, another GnRH (100μg) administration 2 d after PGF2α, and were inseminated 8-18h later (OvSynch). First service conception rate (CR) was determined by rectal palpation at 42 ± 7 d after artificial insemination (AI). Treated cows (n=113) received rbST 67 ± 7 d PP whereas control cows (n = 113) were presynchronized without rbST. The cycling status of all cows was determined by paired milk P4 levels at 53 and 67 ± 7 d PP. No differences (P > 0.10) in conception rate were observed between control and rbST treated cows (44.7 and 40.7% respectively), nor was there any interaction of cyclicity and rbST. Milk samples were collected 7 d following AI for use in ECF test kit evaluation. Samples were stored at -20ºC (n=216) and at 5ºC (n=113) until assayed. Test results for frozen and refrigerated samples were compared to conception rates determined by rectal palpation at 42 ± 7 d after AI. The rate of false positive and negative results for frozen milk samples were 36.1 and 14.8% respectively, and 40.7 and 7.1% for refrigerated milk samples. Treatment with rbST at the time of the first GnRH injection of an OvSynch protocol did not significantly alter first service conception rates. Additionally, an acceptable 92.9% accuracy of the ECF test for the detection of open cows 7 d after AI using milk samples stored at 5ºC was obtained.