Ligation of 5' DNA overhangs to 3' DNA overhangs using short single-stranded oligonucleotide linkers.
نویسندگان
چکیده
17.Summers, A.O. 1992. Untwist and shout: a heavy metal responsive transcriptional regulator. J. Bacteriol. 174:3097-3101. 18.Trudel, P., S. Provost, B. Massie, P. Chartrand and L. Wall. 1996. pGATA: a positive selection vector based on the toxicity of the transcription factor GATA-1 to bacteria. BioTechniques 20:684-693. 19.Vernet, T., P.C.K. Lau, S.A. Narang and L.P. Visentin. 1985. A direct-selection vector derived from pColE3-CA38 and adapted for foreign gene expression. Gene 34:87-93.
منابع مشابه
Presence of double-strand breaks with single-base 3' overhangs in cells undergoing apoptosis but not necrosis
Apoptotic cells in rat thymus were labeled in situ in paraffin-embedded and frozen tissue sections by ligation of double-stranded DNA fragments containing digoxigenin or Texas red. Two forms of double-stranded DNA fragments were prepared using the polymerase chain reaction: one was synthesized using Taq polymerase, which yields products with single-base 3' overhangs, and one using Pfu polymeras...
متن کاملJoining of long double-stranded RNA molecules through controlled overhangs.
We describe two methods for creating long (>1 kb) dsRNA molecules with specific, user-controlled overhangs for efficient hybridization and ligation. The two methods create double-stranded RNA (dsRNA) molecules with 5' overhangs or with 3' overhangs using T7 RNA polymerase (T7 RNAP) in transcription reactions of carefully designed PCR products. Primers utilized in the PCR reactions provide the t...
متن کاملComparative analysis of the end-joining activity of several DNA ligases
DNA ligases catalyze the repair of phosphate backbone breaks in DNA, acting with highest activity on breaks in one strand of duplex DNA. Some DNA ligases have also been observed to ligate two DNA fragments with short complementary overhangs or blunt-ended termini. In this study, several wild-type DNA ligases (phage T3, T4, and T7 DNA ligases, Paramecium bursaria chlorella virus 1 (PBCV1) DNA li...
متن کاملAn efficient method for generation and subcloning of tandemly repeated DNA sequences with defined length, orientation and spacing.
Tandemly repeated DNA sequences generated from single synthetic oligonucleotide monomers are useful for many purposes. With conventional ligation procedures low yields and random orientation of oligomers makes cloning of defined repeated sequences difficult. We solved these problems using 2 bp overhangs to direct orientation and random incorporation of linkers containing restriction sites durin...
متن کاملRapid and efficient cloning of Alu-PCR products using uracil DNA glycosylase.
By incorporating dUMP residues into the 5' end of PCR primers, one can generate products which, after treatment with uracil DNA glycosylase (UDG), contain 3' overhangs. These overhangs can be annealed to vector molecules with complementary overhangs generated in a similar fashion and transformed directly into Escherichia coli without the need for ligase. We have tested this method of ligation-i...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید
ثبت ناماگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید
ورودعنوان ژورنال:
- BioTechniques
دوره 23 3 شماره
صفحات -
تاریخ انتشار 1997