Cloning PCR products into T vectors.
نویسندگان
چکیده
MATERIALS Bacteriophage T4 DNA ligase T vector Target DNA (25 μg/ml), amplified by PCR When the PCR mixture contains more than one or two bands of amplified DNA, purify the target fragment by electrophoresis through low melting/gelling temperature agarose (please see Recovery of DNA from Low-meltingtemperature Agarose Gels: Organic Extraction). If not purified by gel electrophoresis, PCR-amplified DNA should be prepared for ligation by extraction with phenol:chloroform and ultrafiltration through a Centricon-100 filter (please see Removal of Oligonucleotides and Excess dNTPs from Amplified DNA by Ultrafiltration).
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عنوان ژورنال:
- CSH protocols
دوره 2006 1 شماره
صفحات -
تاریخ انتشار 2006