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Identification of transgenic mice by direct PCR analysis of lysates of epithelial cells obtained from the inner surface of the rectum.

作者信息

Lahm H, Hoeflich A, Rieger N, Wanke R, Wolf E

机构信息

Institute of Molecular Animal Breeding, Genecenter, Munich, Germany.

出版信息

Transgenic Res. 1998 Mar;7(2):131-4. doi: 10.1023/a:1008824509988.

DOI:10.1023/a:1008824509988
PMID:9608740
Abstract

Conventional screening protocols for transgene integration in mice employ tail tips or blood samples as sources to obtain genomic DNA preparations. We have developed a simple alternative non-surgical method. Epithelial cells are scraped off the inner surface of the rectum with a sterile plastic inoculation loop and are lysed with Kawasaki buffer. The lysate can be directly examined in a polymerase chain reaction (PCR) analysis without any need for further DNA purification. This procedure causes minimal harm and stress to the animals and repeated samples can be obtained as often as necessary. This technique has been used successfully to identify transgenic mice from a number of different lines. The method allows quick screening of numerous animals and contributes to a reduction of the number of surgical biopsies required.

摘要

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Adverse effects of vapocoolant and topical anesthesia for tail biopsy of preweanling mice.用于断奶前小鼠尾部活检的蒸发冷却液和局部麻醉的不良反应。
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Local cryoanalgesia is effective for tail-tip biopsy in mice.局部冷冻镇痛对小鼠尾尖活检有效。

本文引用的文献

1
Identification of transgenic mice by PCR analysis of saliva.通过唾液的聚合酶链反应(PCR)分析鉴定转基因小鼠。
Nat Biotechnol. 1996 Sep;14(9):1146-8. doi: 10.1038/nbt0996-1146.
2
Actions and interactions of growth hormone and insulin-like growth factor-II: body and organ growth of transgenic mice.生长激素与胰岛素样生长因子-II的作用及相互作用:转基因小鼠的身体和器官生长
Transgenic Res. 1997 May;6(3):213-22. doi: 10.1023/a:1018494108654.
3
Cortisol decreases IGF-I mRNA levels in human osteoblast-like cells.皮质醇可降低人成骨样细胞中IGF-I信使核糖核酸的水平。
J Am Assoc Lab Anim Sci. 2013 Mar;52(2):171-5.
4
Evaluation of tail biopsy collection in laboratory mice (Mus musculus): vertebral ossification, DNA quantity, and acute behavioral responses.实验室小鼠(小家鼠)尾部活检采集的评估:椎体骨化、DNA数量及急性行为反应
J Am Assoc Lab Anim Sci. 2008 Nov;47(6):10-8.
5
A rapid and simple PCR-based method for analysis of transgenic fish using a restricted amount of fin tissue.一种基于聚合酶链反应(PCR)的快速简便方法,用于利用有限量的鳍组织分析转基因鱼。
Transgenic Res. 2003 Aug;12(4):523-5. doi: 10.1023/a:1024274508052.
J Endocrinol. 1996 Jun;149(3):397-403. doi: 10.1677/joe.0.1490397.
4
Genomic DNA analysis from mouse toe lysates.来自小鼠脚趾裂解物的基因组DNA分析。
Transgenic Res. 1996 May;5(3):209-11. doi: 10.1007/BF01969711.
5
Interleukin 4 down-regulates expression of c-kit and autocrine stem cell factor in human colorectal carcinoma cells.白细胞介素4下调人结肠癌细胞中c-kit和自分泌干细胞因子的表达。
Cell Growth Differ. 1995 Sep;6(9):1111-8.
6
Direct PCR from solid tissues without DNA extraction.无需提取DNA即可直接从实体组织进行PCR。
Nucleic Acids Res. 1993 Sep 25;21(19):4656. doi: 10.1093/nar/21.19.4656.
7
A simple method of DNA extraction from whole tissues and blood using glass powder for detection of transgenic animals by PCR.
Transgenic Res. 1995 Mar;4(2):149-50. doi: 10.1007/BF01969417.
8
Consequences of postnatally elevated insulin-like growth factor-II in transgenic mice: endocrine changes and effects on body and organ growth.转基因小鼠出生后胰岛素样生长因子-II升高的后果:内分泌变化以及对身体和器官生长的影响。
Endocrinology. 1994 Nov;135(5):1877-86. doi: 10.1210/endo.135.5.7525257.