Identifying Mutations in Drosophila Genes by Direct Sequencing of PCR Products
نویسندگان
چکیده
منابع مشابه
Direct sequencing of PCR products in agarose gel slices.
Direct DNA sequencing of amplified polymerase chain reaction (PCR) products offers several advantages over cloning of amplified DNA products. It is faster (1 day versus 3—5 days) and in DNA samples containing sequence polymorphisms both the normal and mutated sequence can be detected in the same sequencing reaction. The major problems encountered in direct sequencing of amplified DNA have been ...
متن کاملDirect sequencing of PCR products using unlabeled primers.
An improved protocol is described for using lambda exonuclease to directly sequence PCR products. It is important not to execute PCR cycles beyond the plateau of amplification. The asymmetric PCR and double-stranded DNA sequencing by a snap-cooling procedure were also performed using the same DNA samples and primers. The improved method was the most reliable and produced the best results.
متن کاملRapid sequencing of unpurified PCR products by thermal asymmetric PCR cycle sequencing using unlabeled sequencing primers.
Numerous methods have been developed for direct sequencing of single or double-stranded DNA amplified by PCR, and most require template purification steps prior to sequencing to remove excess unincorporated primers and dNTPs. Purification methods can be cumbersome, laborious and time consuming, or yield low DNA recoveries. Strategies for direct sequencing of doublestranded DNA products without ...
متن کاملIdentifying insertion mutations by whole-genome sequencing.
Insertion mutagenesis via mobile genetic element is a common technique for the analysis of gene function in model organisms. Next-generation sequencing offers an attractive approach for localizing the site of insertion, but alignment-based mapping of mobile genetic elements is challenging. A computational method for identifying insertion sites is reported herein. The technique was validated by ...
متن کامل[ABO genotyping by PCR-direct sequencing method].
OBJECTIVE To analyze the sequence difference between human A, B, and O alleles and establish the method of ABO genotyping by PCR direct sequencing. METHODS PCR-direct sequencing technique was used to analyze two regions of cDNA from A transferase gene, 233-433 and 660-788. RESULTS Two nucleotide substitutions at 258th and 297th were found in 233-433 region, and a nucleotide substitution at ...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
ژورنال
عنوان ژورنال: BioTechniques
سال: 1999
ISSN: 0736-6205,1940-9818
DOI: 10.2144/99272bm09