基因標(biāo)的載體建構(gòu)服務(wù)使用規(guī)范

1、A4-1 基因標(biāo)的載體建構(gòu)服務(wù)使用規(guī)範(fàn)基因轉(zhuǎn)殖小鼠模型核心設(shè)施(TMMC)基因標(biāo)的載體建構(gòu)服務(wù)使用規(guī)範(fàn)1. 使用者需提供本核心建構(gòu)標(biāo)的載體所需的基因體DNA片段，核心提供含有抗生素篩選基因的標(biāo)的骨架。2. 基因標(biāo)的載體使用的基因體DNA片段須與本核心設(shè)施使用的胚胎幹細(xì)胞株(R1：129/SvJ 129/Sv-CP；E14：129/Ola)同源，以確保較高的標(biāo)的機(jī)率。使用者如提出要求使用C57BL/6胚胎幹細(xì)胞，則須經(jīng)使用者委員會審核裁示。3. 本核心構(gòu)築標(biāo)的載體的方法是採用重組工程法(Recombineering)a，該方法乃分別使用Neo (Neomycin phosphotransfera

2、se)和胸腺嘧啶激酶(Thymidine kinase)為正負(fù)篩選標(biāo)記。4. 使用者送件前需填寫本核心網(wǎng)站提供之基因標(biāo)的載體建構(gòu)服務(wù)申請書及同意書並簽名繳交。本核心於收件時(shí)將與使用者共同填寫使用同意證明單（四聯(lián)單），使用者憑此四聯(lián)單繳費(fèi)。 5. 每次基因標(biāo)的載體建構(gòu)服務(wù)價(jià)格：學(xué)術(shù)單位 ：NT25,000元 非學(xué)術(shù)單位:：NT35,000元a: A Highly Efficient Recombineering-Based Method for Generating Conditional Knockout Mutations. Pentao Liu, Nancy A. Jenkins, and

3、 Neal G. Copeland. Genome Res. 2003,13:476-84.Targeting Construct Production Request Form (A4-1.1) p1Transgenic Mouse Model Core FacilityTargeting Construct Production Request FormDate： / / 20 Print P.I. nameDepartment/ InstituteTelephone numberFAX numberE-mail addressP.I. addressGrant nameGrant sou

4、rce/ grant numberContact personTelephone number/FAX numberE-mail addressSignature of P.I.Construct name： Construct description (skip if unknown or unapplicable)Nature of construct：knock-out knock-in conditional others_Knock-in origin (please specify any possible correlation to disease)species：mouse

5、human others_Gene function： Phenotypes： Disease-correlation：No Yes, please notify the disease： _Targeting Construct Production Service Request Form (A4-1.1) p2Detailed construct mapPlease notify the position, size and orientation of the promoter, the insert, multiple cloning sites, antibiotic used f

6、or selection, fragment used for microinjection, restriction enzymes used to release the transgene fragment.*record number of the target gene construct：# _* scheduled date： / /200 ( * Filled by TMMC only)Targeting Construct Production Service Agreement Form (A4-1.2)Transgenic Mouse Model Core Facilit

7、yTargeting construct production Service Agreement FormAll TMMC services require the agreement and signature from service user with full understanding of all the following statements:1. Transgenic mouse model core facility (TMMC) will provide high quality service of targeting construct production, al

8、l requests are treated confidentially.2. TMMC provide technique service for targeting construct production only. TMMC will not take responsibility for any unexpected results or damage caused by targeting construct.3. Requester P.I. should obtain approval of the original designer of the gene construc

9、t should the request construct has been used for another transgenic mouse model before. 4. TMMC provide technique service for targeting construct production only. Expression of phenotypes is not guaranteed.5. Any productive outcome, such as publications and patents, should acknowledge the TMMC of NR

10、PGM.All TMMC services require the agreement and signature from service user with full understanding of all the following statements:a. I have carefully reviewed the TMMC guideline and condition for using the service, and I agree to follow completely to the TMMC guideline.b. I acknowledge that TMMC reserves rights to reject or stop my servi