DNA电泳

DNA电泳（主要内容如下）

•   Preparation of Agarose Gel and Electrophoresis

•   Extraction of DNA From Agarose Gel

•   Extraction of DNA from Acrylamide Gels

•   DNA Marker

•   Denaturing Gradient Gel Electrophoresis

•   Two Dimensional Gel Electrophoresis

•   DNA Visualization and Staining

•   In Gel Application

•   Migration Table

•   Q&A posted in the method forum

Preparation of Agarose Gel and Electrophoresis

·         Top 10 Fun Facts for DNA Electrophoresis (Produced from LTI)
Useful tips for preparing agarose gel and electrophoresis

·         Agarose Gel Electrophoresis (LaboratoryExperiments.com)
A basic and very useful protocol for gel preparation and electrophoresis. 

·         Agarose Gel (John Garland)
How to prepare stock solutions and agarose solution

·         Dissolving and Casting Agarose Gels (FMC)
Very nice and detailed procedures for preparing gel and some basic knowledge about agarose gel

·         Agarose Gel Electrophoresis (Molecular Biology Cyberlab)
Very basic and illustrated guide  

·         Agarose Gel Electrophoresis (Julie B. Wolf, UMBC)
Provides method in gel preparation, recipes of electrophoresis buffer and loading buffer...

·         Agarose Gel Electrophoresis (Frank Bottone, Jr.)

·         DNA Gel (Hoshi Lab)
Offers detailed information about resolution of DNA with agarose gel, gel preparation and electrophoresis...

·         Agarose Electrophoresis (Chen)
DNA electrophoresis, extraction and concentration determination.

·         Agarose Gel Electrophoresis (Roe, OU)

·         DNA Mobility in Gels (Roe, OU)

·         Trouble Shooting DNA Agarose Gels (LTI)
No band, Smear, anomalies band migration? Find trouble shooting here

• DNA Electrophoresis in Agarose Gel (FMC)
  General and useful tip for agarose gel electrophoresis including the following
  Buffers for Electrophoresis  
  Suggested Agarose Concentrations  
  Dye Mobility Tables  
  Running Conditions for Agarose Gels
  DNA Loading and Loading Buffers

Extraction of DNA From Agarose Gel

• Tips for Increasing DNA Recovery Efficiency from Agarose Gels (FMC)
    

• ß-Agarase Recovery of DNA from Agarose Gels (FMC)
    

• Electroelution of DNA from Agarose Gels (FMC)
    

• Phenol/Chloroform Extraction of DNA from Agarose Gels (FMC)
    

• "Modified Freeze/Squeeze" Extraction of DNA from Agarose Gels (FMC)
    

• DNA Extraction from Agarose Gel (LaboratoryExperiments.com)
  Protocol for extracting DNA from agarose gel using fileter.
    

• Ethanol Precipitation of DNA Recovered from Agarose Gels (FMC)
    

• Other Recovery Methods from Agarose Gels (FMC)

·         Elution of DNA fragments from agarose (Roe, OU)

·         DNA Extraction from Agarose Gel (Crawford Lab)

·         Beta-Agarase Gel Purification  (Crawford Lab)
Recovering DNA from Agarose gel using Beta-Agarase

• Quick Isolation of DNA From Agarose Gel (Hancock Lab) (Accessible only by Internet Explorer)
  Centrifugation and subsequent phenol extraction method
    

• Isolation of DNA fragments from agarose gels. (Freeze squeeze) (Molecular Genetics Rec Lab)
    

• Isolation of DNA fragments from agarose gels. (Freeze spin) (Molecular Genetics Rec Lab)

Extraction of DNA from Acrylamide Gels

·         Isolation of DNA from Acrylamide Gels (Schimenti Lab)

·         Elution of DNA Fragments from Polyacrylamide Gels (Molecular Genetics Rec Lab)

Molecular Marker

• FAQs for Molecular Weight Standards and Nucleic Acids (LTI)
  So many FAQs about molecular weight standards, such as what can I do to prevent DNA standard from smearing? How can I quantitate DNA on an agarose gel? How can I label DNA ladder and more...

·         DNA Fragment Length Standards (Molecular Genetics RecLab)
Lists of DNA restriction fragment length of lambda and pBR322

DGGE

·         Denaturing Gradient Gel Electrophoresis (DGGE) (Donis-Keller lab)
Denaturing gradient gels are used to detect non-RFLP polymorphisms. The small (200-700 bp) genomic restriction fragments are run on a low to high denaturant gradient acrylamide gel; initially the fragments move according to molecular weight, but as they progress into higher denaturing conditions, each (depending on its sequence composition) reaches a point where the DNA begins to melt. The partial melting severely retards the progress of the molecule in the gel, and a mobility shift is observed. It is the mobility shift which can differ for slightly different sequences (depending on the sequence, as little as a single bp change can cause a mobility shift). Alleles are detected by differences in mobility.

Two Dimensional Gel Electrophoresis

• Standard 2-D Gel of DNA Replication Intermediatess (Fangman/Brewer)
  The first dimension gel is intentionally run at low voltage in low percentage agarose to separate DNA molecules in proportion to their mass. The second dimension is run at high voltage in a gel of higher agarose concentration in the presence of Ethidium bromide so that the mobility of a non-linear molecule is drastically influenced by its shape.
    

• Fork-Direction 2-D Gels (Fangman/Brewer)
  For detecting direction of replication fork movement 

DNA Staining

• Visualizing DNA in Agarose Gels (FMC)
  Provides several detection methods: SYBR^® Green I staining, ethidium bromide staining, silver staining and acridine orange and methylene blue staining.

·         Methylene Blue DNA staining (Gerard Lazo)
This method primarily eliminates the damage of DNA by uv irradiation. DNA isolated from methylaene blue stained gels should transform frozen competent E. coli cells on the order of 20-50 fold more efficiently than ethdium bromide isolated DNA.

·         Vacuum Drying Agarose Gels (FMC)
Many people ask how to dry agarose gel, here is the answer

In Gel Application

A usr/localiety of enzymatic reactions can be performed in the presence of agarose. In-gel reactions are an alternative approach to standard DNA recovery techniques and provide a multitude of benefits. The use of in-gel reactions will not only save time, but eliminate potential sample loss during DNA recovery from an agarose gel (FMC)

• Tips for Increasing the Efficiency of In-Gel Reactions (FMC)
    

• Random-Prime Labeling in the Presence of Agarose (FMC)
    

• Cloning in the Presence of Agarose (FMC)
    

• In-Gel Cloning (Crawford Lab)
  Digest your vector and insert DNA and purify by gel electrophoresis and do ligation without further recovering DNA from gel.  It works great!
    

• Restriction Digestion in the Presence of Agarose (FMC)
    

• DNA Amplification in the Presence of Agarose (FMC)
    

• Core sample PCR (NWFSC)
  A method to re-PCR unique bands from products of mixed size