Direct sequencing of PCR products in agarose ge l slices
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 کاملDirect single stranded sequencing from agarose of polymerase chain reaction products.
The polymerase chain reaction (PCR) has made possible the rapid isolation and amplification of specific DNA segments, which can then be used for a wide range of applications (1). Direct sequencing of PCR products has been described, using assymetric PCR, which requires purification of the PCR product, or inclusion of 10% DMSO in the sequencing reaction (2, 3, 4). However, these methods of seque...
متن کاملPreparation of PCR products for DNA sequencing.
We demonstrate that routine PCR product analytical agarose gels can also serve as preparative gels for quick DNA template purification before sequencing. The band of interest is excised, placed into a Gel Nebulizer inside a Micropure separator and rapidly purified in a single centrifugation step. Gel-purified PCR product, suitable for manual and automated sequencing, is delivered within 10 min.
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 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: Nucleic Acids Research
سال: 1994
ISSN: 0305-1048,1362-4962
DOI: 10.1093/nar/22.16.3425