Feather is a noninvasive material for avian molecular genetic analysis.Calamus has been proved to be a good source of DNA and widely used.Vane and rachis is also a source of DNA,although the DNA is significantly lower in both quality and quantity.DNA from the calamus is not always available for molecular genetic studies,and so the usability of vane and rachis DNA needs to be evaluated.By using polymerase chain reaction (PCR) amplification of variably-sized fragments of the cytochrome c oxidase I (COI) gene based on DNA isolated from remiges and contour feathers of four passerine birds,we evaluated the usability of vane and rachis,including (1) the influences of purification on the success rate of PCR amplification; (2) the influence of different parts of remiges on the success rate of PCR amplification; and (3) the size affordability of DNA recovered from different part of remiges and whole contour feather for PCR amplification.Results showed that the purification process was able to significantly improve the success rate relative to the routine phenol-chloroform method.The calamus of both remiges and contour feather could afford up to 736 bp amplicons,while vane and rachis of remiges could afford <354 bp fragments.By using a purification approach,the success rate was 75.0% to 100% for small (<354 bp) amplicons,but 0 for large (746 bp) amplicons in distal remiges (vane+rachis).The success approached 83.3% to 100% for amplicons <354 bp,and 50% for 746 bp amplicons in proximal parts.The success rate for whole contour feathers was high for the whole size range of amplicons due to the presence of calamus.