Straightforward Procedure for Laboratory Production of DNA Ladder

نویسندگان

  • Vo Thi Thuong Lan
  • Pham Thi Thanh Loan
  • Pham Anh Thuy Duong
  • Le Thi Thanh
  • Ngo Thi Ha
  • Ta Bich Thuan
چکیده

DNA ladder is commonly used to determine the size of DNA fragments by electrophoresis in routine molecular biology laboratories. In this study, we report a new procedure to prepare a DNA ladder that consists of 10 fragments from 100 to 1000 bp. This protocol is a combination of routinely employed methods: cloning, PCR, and partial digestion with restriction enzymes. DNA fragments of 100 bp with unique restriction site at both ends were self-ligated to create a tandem repeat. Once being cloned, the tandem repeat was rapidly amplified by PCR and partially digested by restriction enzymes to produce a ladder containing multimers of the repeated DNA fragments. Our procedure for production of DNA ladder could be simple, time saving, and inexpensive in comparison with current ones widely used in most laboratories.

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

Simple procedure for production of short DNA size markers of 100 to 2000 bp

DNA size markers (ladder) are essential tools in molecular biology, genetics and biotechnology. In this study, a simple and cost-effective method for laboratory production of DNA ladders is introduced. For this purpose, different sizes of 100 to 2000 bp DNA segments were designed using PCR technique. For producing 14 different gene fragments as DNA molecular weight markers, recombinant plasmid ...

متن کامل

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

LAMP assay for rapid diagnosis of cow DNA in goat milk and meat samples

Animal species detection is one of the crucial steps for consumer’s food analysis. In the present study we developed an in-house built loop-mediated isothermal amplification (LAMP) assay for rapid detection of adulterated cow DNA in goat milk/meat samples. The cow milk/tissue DNA in goat milk/meat samples were identified in the developed LAMP assay by either naked eye visualizing with SYBR Gree...

متن کامل

DNA Polymorphisms at Candidate Gene Loci and Their Relation with Milk Production Traits in Murrah Buffalo (Bubalus bubalis)

DNA polymorphism within diacylglycerol transferase 2 (DGAT2) / monoacyl glycerol transferases 2 (MOGAT2), leptin and butyrophilin genes were analysed using PCR-SSCP in Murrah buffalo. The single strand conformation polymorphism (SSCP) analysis of amplified gene fragment in exon 5 of MOGAT2, exon 3 of leptin and intron 1 of butyrophilin gene revealed different patterns. A, B and C showed the fol...

متن کامل

A Simple and Effective Protocol for One-Pot Diazotization-Iodination of Aromatic Amines by Acidic Ionic Liquid [H-NMP]HSO4 at Room Temperature

A simple and efficient one-pot method for the preparation of aromatic iodides has been developed by sequential diazotization–iodination of aromatic amines employing sodium nitrite and sodium iodide in the presence of acidic ionic liquid N-methyl-2-pyrrolidonum hydrosulfate ([H-NMP]HSO4). The diazonium salts that are formed by this ionic liquid are stable at room temperature and r...

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:

دوره 2012  شماره 

صفحات  -

تاریخ انتشار 2012