Preparation of PCR products for DNA sequencing.

نویسندگان

  • J T Leonard
  • M B Grace
  • G S Buzard
  • M J Mullen
  • C B Barbagallo
چکیده

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.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

Construction of a Synthetic Vector for Preparation of a 100 Base Pair DNA Ladder

DNA size markers are widely used to estimate the size of DNA samples on agarose or polyacrylamide gelelectrophoresis (PAGE). DNA markers can be prepared by mixing PCR products with definite sizes.Alternatively, they are prepared by restriction enzyme digestion of the genomic DNA of bacteriophages ornatural and synthetic DNA plasmids. The present study describes engineering of ...

متن کامل

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 کامل

Reflex™: a novel method to sequence barcoded long-range PCR products in a pooled population of hundreds of DNA samples

We are developing novel methods to simultaneously analyze candidate genes or regions from multiple samples in a single experiment rapidly and cost-effectively. One method, we call Reflex, provides an elegant approach to sample preparation for long-range PCR (LR-PCR) amplicons. Current methods use LR-PCR for target enrichment then use random fragmentation of each sample followed by ligation of D...

متن کامل

DNA sequencing with Thermus aquaticus DNA polymerase and direct sequencing of polymerase chain reaction-amplified DNA.

The highly thermostable DNA polymerase from Thermus aquaticus (Taq) is ideal for both manual and automated DNA sequencing because it is fast, highly processive, has little or no 3'-exonuclease activity, and is active over a broad range of temperatures. Sequencing protocols are presented that produce readable extension products greater than 1000 bases having uniform band intensities. A combinati...

متن کامل

Sequencing of genomic DNA by combined amplification and cycle sequencing reaction.

BACKGROUND Despite considerable advances, DNA sequencing has remained somewhat time-consuming and expensive, requiring three separate steps to generate sequencing products from a template: amplification of the target sequence; purification of the amplified product; and a sequencing reaction. Our aim was to develop a method to routinely combine PCR amplification and cycle sequencing into one sin...

متن کامل

ذخیره 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 نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:
  • BioTechniques

دوره 24 2  شماره 

صفحات  -

تاریخ انتشار 1998