CHEF Genomic DNA Plug Kits

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Overview

Introduction to Pulsed Field Gel Electrophoresis
Introduction to Pulsed Field Gel Electrophoresis

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Separation of E. coli
Separation of E. coli Chromosomal Restriction Fragments

Separation of E. coli chromosomal restriction fragments. Lanes 1 and 14, S. cerevisiae chromosomes; lanes 2 and 13, lambda ladder; lanes 3–6, NotI restriction digests of chromosomal DNA from DH5αF' (lane 3), HB101 (lane 4), MC1061 (lane 5), MV1190 (lane 6); lanes 8–11, SfiI restriction digests of chromosomal DNA from DH5αF' (lane 8), HB101 (lane 9), MC1061 (lane 10), MV1190 (lane 11). DNA fragments were separated on a 1.0% pulsed field certified agarose gel in 0.5x TBE at 14°C for 26.4 hr at 6 V/cm (200 V) using a 120° included angle with a 2.77–26.32 sec linear switch-time ramp.
PFGE Standards Selection Guide
PFGE Standards Selection Guide
5 Mb Ladder 8–48 kb Ladder Lambda Ladder S. cerevisiae
Chromosomal DNA		 H. wingei
Chromosomal DNA		 S. pombe
Chromosomal DNA
Comparison of Pulsed Field Standards
 
  Range Contents Amount Number of Applications
 
5 kb ladder 4.9–120 kb Concatemers of pBR328 20 µg in 200 µl 20–25
8–48 kb ladder 8.3–48.5 kb Mixed digest of phage λ 25 µg in 125 µl 125
Lambda (λ) ladder 0.05–1 Mb Concatemers of phage λcl857Sam7 5 agarose blocks 25–40
S. cerevisiae 0.225–2.2 Mb Saccharomyces cerevisiae chromosomal DNA 5 agarose blocks 25–40
H. wingei 1–3.1 Mb Hansenula wingei chromosomal DNA 5 agarose blocks 25–40
S. pombe 3.5–5.7 Mb Schizosaccharomyces pombe chromosomal DNA 5 agarose blocks 25–40
 

 

Certified Agarose Selection Guide
Certified Agarose Selection Guide
 
Packaging
25 g 161-3100 161-3103
 161-3106   161-3111 161-3113 162-0137
100 g             162-0138
125 g 161-3101 161-3104 161-3107   161-3112 161-3114  
500 g 161-3102 161-3105       161-3115  
Analytical Separation
≥1,000 bp •       •    
≤1,000 bp   •       •  
20–800 bp     •        
1 kb–2 Mb       •     •
1 kb–5 Mb       •      
Preparative Separation
Quantum Prep methods • • • •      
Low-melt methods         • •  
Agarose methods         • •  
In-gel applications         • •  
Other Applications
Postpreparative enzymatic treatments • • •   • •  
Tissue/cell culture       •      
Pulsed field sample preparation       •     •
Blotting • •   •     •

CHEF genomic DNA plug kits provide a convenient means for preparing intact, chromosome-sized DNA for PFGE. With such large molecules, pipetting mechanically shears the DNA, resulting in unacceptable quality for PFGE separations. This problem has required procedures for lysis and deproteinization of whole cells embedded in agarose to yield intact purified DNA. Three kits are available for the preparation of bacterial (lysozyme-sensitive) or mammalian genomic DNA and yeast chromosomes (YACs). Each kit contains all the enzymes, reaction buffers, and restriction digest-qualified CleanCut agarose necessary to prepare 100 plugs, as well as disposable plug molds and screened caps for simplified plug processing. Each kit is thoroughly tested to ensure that prepared genomic DNA can be restriction digested and separated on a CHEF electrophoresis system.

CHEF Mammalian Genomic DNA Plug Kit  
Cell suspension buffer 12 ml     
>600 U/ml proteinase K 1.3 ml
Proteinase K reaction buffer 30 ml
2% CleanCut™ agarose 12 ml
10x wash buffer 60 ml
50-well disposable plug mold 2
Screened cap 1
CHEF Bacterial Genomic DNA Plug Kit  
Cell suspension buffer  12 ml 
>600 U/ml proteinase K  1.3 ml  
Proteinase K reaction buffer  30 ml 
2% CleanCut agarose  12 ml    
10x wash buffer  60 ml  
25 mg/ml lysozyme 1.6 ml     
Lysozyme buffer 30 ml  
50-well disposable plug mold  2 
Screened cap  1 
CHEF Yeast Genomic DNA Plug Kit  
Cell suspension buffer   12 ml  
>600 U/ml proteinase K      1.3 ml  
Proteinase K reaction buffer   30 ml  
2% CleanCut agarose   12 ml    
10x wash buffer   60 ml    
5,000 U/ml lyticase 1.6 ml   
Lyticase buffer 30 ml    
50-well disposable plug mold     2    
Screened cap 1    
CHEF Mammalian Genomic DNA Plug Kit

170-3591
Mammalian preparation kit for PFGE, includes proteinase K, CleanCut Agarose, lyticase, all required buffers, screened cap, 2 plug molds, for 100 plugs

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CHEF Bacterial Genomic DNA Plug Kit

170-3592
Bacteria preparation kit for PFGE, includes proteinase K, CleanCut Agarose, lysozyme, required buffers, screened cap, 2 plug molds, for 100 plugs

List Price:   $415.00
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CHEF Yeast Genomic DNA Plug Kit

170-3593
Yeast preparation kit for PFGE, includes proteinase K, CleanCut Agarose, lyticase, all required buffers, screened cap, and 2 plug molds, for 100 plugs

List Price:   $480.00
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CleanCut Agarose

170-3594
12 ml, 2% agarose for pulsed field gel electrophoresis (PFGE), restriction digest-qualified, for 100 plugs

List Price:   $30.00
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Number Description Options
1753 Preparation and Restriction Digestion of Escherichia coli Chromosomal DNA in Agarose Plugs for Use in PFGE Bulletin, Rev A Click to download
LIT510 Instruction Manual, CHEF Genomic DNA Plug Kit, Rev E Click to download
5901 Pulsed Field Gel Electrophoresis Brochure, Rev B Click to download [ Add to Cart ]