DNA抽提

DNA抽提（主要内容如下）

·   Working with DNA

·   DNA Extraction from Bacteria and Other Organisms

·   DNA Extraction from Cell and Tissue

·   Mitochondria DNA Isolation

·   DNA Extraction from Plants, Fly, Fungal

·   DNA Precipitation and Purification

·   DNA Extraction from Agarose Gel

·   DNA Extraction from Acrylamide Gel

·   Q&A posted in the method forum

Working with DNA

·         Working with DNA (Julie B. Wolf, UMBC)
DNA storage, concentration, purification, quantification

·         Working with DNA and RNA  (Ambion)
Tips on handing nuclear acid during phenol extraction, ethanol precipitation.

DNA Extraction from Bacteria and Other Organisms (see also Bacteria> DNA Extraction) 

·         DNA Extraction from Bacteria (Julie B. Wolf,UMBC)
Phenol/chloroform method

·         DNA Extraction From Bacteria (Triton Method) (NWFSC)

·         Rapid genomic DNA isolation from P. aeruginosa (Goldberg Lab)

·         DNA Extraction from Gram-positive Bacteria (NWFSC)
This procedure was originally developed for Listeria monocytogenes but has worked well with other Gram+ bacteria

• Extraction of Total Genomic DNA from Single Lice, Mites or Other Terrestrial Microarthropods (Robert H. Cruickshank)
  efficient and quick glassmilk method for extracting DNA
    

• Alternative Method for Extraction of Total Genomic DNA from Single Lice, Mites or Other Terrestrial Microarthropods  (Robert H. Cruickshank)

DNA Extraction from Cell and Tissue

·         DNA Extraction from Tissue (Crawford Lab)

·         Simplified DNA extraction from cell or tissue (Laird PW)
This method doesn't require organic extraction and centrifugation. It's the most simplest way of preparing DNA and works great. 

• DNA Extraction from Mouse Liver (Chastain)
  Very basic protocol for students

• Genomic DNA Isolation from Blood (Roe Lab)
  Standard protocol for DNA isolation from blood
    

• Microdissection and DNA Extraction from Paraffin Sections (Waldman Lab)
  Collecting tissue from paraffin section for DNA extraction using microdissection technique.
     

• Extraction of DNA from Paraffin Sections without Microdissection (Waldman Lab)

• Manual Isolation of Human DNA from Lymphoblasts or Whole Blood (Donis Keller Lab)
  Nucleated blood cells are used to prepare genomic DNA. The cells are first washed, then lysed and the nuclei (left intact) are pelleted in a low speed centrifuge run. 
       

• Salting Out Procedure for Human DNA Extraction (Donis Keller Lab)
  This procedure avoids using phenol and chloroform by using high salt concentrations to remove proteins. It is rapid, safe and inexpensive. 
      

·         Tomcod DNA Isolation Method (Crawford Lab)

• Genomic DNA Quickprep for PCR (Kay Schneitz Lab)
  Very simple method of preparing DNA for PCR with recipes
     

• Extraction of DNA from the Whole Blood by Silica Gel (Institute of Gene Biology, RAS)
  Simple and inexpensive procedure in which the pure DNA is extracted from the whole blood within 15-20 min. 
    

• Extraction of DNA from Blood for PCR (Jackson Labs)
  Simple method for preparing DNA from blood for PCR without organic extraction
    

• DNA Isolation from Murine Tails without Phenol Chloroform (Immunology Resource)
  Detailed protocol and recipes

• Non-organic Isolation of Tail DNA (Jackson Labs)
  Using tail tissue directly for PCR after proteinase K digestion
    

• DNA Extraction from Tail Biopsies (Jackson Labs)
  Phenol:chloroform extraction method
     

• Isolation of DNA from Mouse Tail Biopsies (University of Michigan Transgenic Animal Model Core)
     

• Preparation of Mouse Tail DNA for Dot Blots or PCR  (Eric Mercer)
  These procedures were originally devised in Richard Palmiter's lab for use with tail dots (DNA spotted onto a nitrocellulose filter and probed for a transgene; quicker than doing southerns), and subsequently modified for PCR preps.
    

• Nucleic Acid Isolation and Purification Manual (BMB)
  This 180-page manual collects a wide usr/localiety of reliable protocols and technical tips and will enable readers to quickly find the most optimal method for their specific needs.
    

·         Solid-Phase Reversible Immobilization (SPRI) Technique for DNA Purification(Whitehead Institute/MIT Center for Genome Research)
SPRI uses the inexpensive carboxyl-coated magnetic particles that comprise the base material for most magnetic particle manufacture. Under conditions of high polyethylene glycol and salt concentration, the surface of these particles binds both single- and double-stranded DNA, including sequencing reaction products, PCR products, M13 phage, lambda phage, plasmids, cosmids and BACs. 

o        Introduction to Solid-Phase Reversible Immobilization (SPRI)

o        SPRI for M13 Clones (mega-high thoughput!)

o        SPRI for PCR products
This solid-phase procedure is fast, simple and highly automatable

o        SPRI For Extension Product Purification

o        M13 Purifications using Streptavidin Coated Magnetic Particles

• M13 DNA Isolation using SPRI
  The SPRI M13 procedure is based upon the solid-phase reversible immobilization techniques developed for double stranded DNA isolation. 
    

·         Phenol/Chloroform Extraction of DNA (UMBC)

·         Isolation of Mouse Genomic DNA (PMCI Research)

·         Phenol extraction of DNA  (Roe Lab)

• De-Paraffinization of Tissue Sections (Arcturus)
  Remove paraffin from section for tissue preparation such as nucleic acid extraction.
     

• DNA Extraction From Paraffin Section (Arcturus)
  This extraction method has been used for measuring loss of heterozygosity (LOH), dideoxy fingerprinting (DDF), clonality analysis (chromosome X inactivation) and direct sequencing of PCR* products for single base mutational analysis. Extractions are typically performed on 500-1000 captured cells.

Mitochondria DNA Isolation

·         Mitochondrial DNA Isolation from Somatic Embryogenic Cell Cultures of Larix(Kim Marshall)

DNA Extraction from Plants, Fly, Fungal

• Preparation of Megabase-size DNA (Clemson University Genomics Institute)
  To construct large insert DNA libraries in BAC and YAC vectors, methods must be developed to isolate very high molecular weight DNA - megabase-size DNA
    

• Isolation of High Molecular Weight Plant DNA (Long Method) (Gimila's Lab)
  A method for the isolation of intact, high molecular weight (>150kbp) plant nuclear DNA for construction of a genomic library
    

• A Rapid Method to Isolate Plant Genomic DNA.(Atle M. Bones)
    

• Phenol/SDS Method for Plant RNA Preparation (Gimila's Lab) 
  
     

• Isolation of Genomic DNA From Corn (Dr. Chastain)
  Very basic protocol for students.
    

• Genomic Plant DNA Extraction for Mapping (Kay Schneitz Lab)
  This protocol is quick, reliable and the DNA obtained can be used for PCR-based mapping using SSLP and CAPS markers.
      

• Isolation of Plant Geneomic DNA with CTAB (Leaf Disc)(Graham Casey)
    

• Rapid DNA Extraction and Purification(Graham Casey)
    

• Half Seed DNA Extraction Protocol(Graham Casey)
    

• Mini DNA extraction from wheat leaves for PCR (Plant Sciences, Montana Univ)
    

• Large Scale Monocot DNA Isolation(Graham Casey)
    

• Seed Extraction Protocol(Graham Casey)
   

• Simultaneous Extraction of Chloroplast and Nuclear DNA  (Szmidt)
     

• Streamlined DNA Extraction Protocol (TGERC)
  This procedure has been tested with a usr/localiety of Populus species, as well as tobacco and Arabidopsis.The resulting DNA is of sufficiently high quality for PCR (including RAPD), restriction digests, and ligation reactions.
    

• DNA Purification-Modified CTAB Procedure (Kim Marshall)
  DNA extraction from needles 
    

• Single-Fly DNA Preparations for PCR (Lazo Lab)
  A simple method for the rapid and reproducible isolation of DNA from single flies for amplification by PCR and direct sequencing by asymmetric PCR.  
    

• Single-Fly DNA Preparations for PCR (UW Laboratory of Genetics)
    

·         Fungal Genomic DNA Extraction (Contributed by Dr. Eric W. Boehm)
This procedure does not require phenol extraction. The DNA is pure enough for restriction digests, PCR and genomic library construction.
   

• DNA Isolation from Fungal (Lazo Lab)

DNA Precipitation and Purification

·         General Protocol for Precipitation of DNA with Sodium Acetate and Ethanol(Dr. Chastain)
Very basic knowledge and protocol for precipitating DNA

·         Ethanol Precipitation of DNA (UMBC)

·         DNA/RNA precipitation (GSC, WU)
Notes on precipitation of nucleic acids

·         Concentration of DNA by ethanol precipitation (Bruce A. Roe)

·         Ethanol Precipitation of DNA (Life Technologies)
Basic facts about DNA precipitation

·         DNA Precipitation (Gerard Lazo)

·         Fragment purification on Sephacryl S-500 spin columns (Bruce A. Roe)
DNA fragments larger than a few hundred base pairs can be separated from smaller fragments by chromatography on a size exclusion column such as Sephacryl S-500.

DNA Extraction from Agarose Gel

• DNA Fragment Purification from Agarose or Acrylamide (Mike A. Dyer)
  For fragments from 200 bp to 10 kb the agarose purification is ideal. For smaller fragments (20 bp to 400 bp) the acrylamide purification is preferred.
    

• Electroelution of DNA Fragments From Agarose into Dialysis Tubing (Donis Keller Lab)
  To retrieve and purify any specific DNA fragment from an agarose gel slice; the expected yield is from 50-75% of the amount in the gel slice.
    

• Isolation of Restriction Fragments from Agarose Gels by Collection onto DEAE Cellulose(Donis Keller Lab)
  Recovery of DNA from agarose gel
    

·         Alternate Method for Purifying DNA From Agarose Gels

·         DNA Isolating From Low Melting-Temperature Agarose (NWFSC)
Phenol-based Method for the Isolation of DNA Fragments from Low-Melting Temperature Agarose

• Elution of DNA fragments from agarose (Roe Lab)
  DNA fragments are eluted from low-melting temperature agarose gels using an unpublished procedure first developed by Dr. Roe. Here, the band of interest is excised, frozen at -70C, and then melted...
     

·         Glass_Powder Purification of_DNA from Agarose Gel(pdf) (Templenton Lab)
Tried and true method for eluting DNA from agarose gels. Like Geneclean, but nearly free.

• DNA Isolation from Agarose Gels with DEAE Paper  (Hancock Lab)
  Detailed protocol with recipes
    

·         DNA Extraction from Agarose Gel (Crawford Lab)
Extract DNA from low melting gel

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

·         DNA Extraction from Agarose  (Hahn Lab)
Glass wool spin column method 
  

·         DNA Isolation from Agarose Gel Using NaI (Corbett Lab)

DNA Extraction from Acrylamide Gel

• DNA Fragment Purification from Agarose or Acrylamide (Mike A. Dyer)
  For fragments from 200 bp to 10 kb the agarose purification is ideal. For smaller fragments (20 bp to 400 bp) the acrylamide purification is preferred.