CHEF Genomic DNA Plug Kits

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CHEF Genomic DNA Plug Kits

Overview

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
5 kb 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
 
Packaging
25 g 1613100 1613103 1613106  1613108 1613111 1613113 1620137
100 g             1620138
125 g 1613101 1613107  1613109 1613112 1613114  
500 g 1613102 1613105    1613110   1613115  
Analytical Separation
≥1,000 bp          
≤1,000 bp          
10–800 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.
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  • Description
  • Specifications
  • Ordering
  • Systems
  • Documents

Description

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.


Specifications

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 
Screened cap 
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    

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CHEF Mammalian Genomic DNA Plug Kit

1703591
Mammalian preparation kit for PFGE, includes proteinase K, CleanCut Agarose, all required buffers, screened cap, 2 plug molds, for 100 plugs

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$445.00 
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CHEF Bacterial Genomic DNA Plug Kit

1703592
Bacteria preparation kit for PFGE, includes proteinase K, CleanCut Agarose, lysozyme, required buffers, screened cap, 2 plug molds, for 100 plugs

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$578.00 
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CHEF Yeast Genomic DNA Plug Kit

1703593
Yeast preparation kit for PFGE, includes proteinase K, CleanCut Agarose, lyticase, all required buffers, screened cap, and 2 plug molds, for 100 plugs

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$662.00 
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CleanCut Agarose

1703594
12 ml, 2% agarose for pulsed field gel electrophoresis (PFGE), restriction digest-qualified, for 100 plugs

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$37.00 
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Systems

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list price. If the problem persists, please call customer service at or send an
email to

CHEF Mapper XA Chiller System, 120 V

1703670
120 V, complete PFGE system, includes cooling module, pump, tubing, S. cerevisiae DNA size standard, 5 g pulsed field Certified agarose, 5 g pulsed field Certified megabase agarose

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CHEF-DR III Chiller System

1703700
120 V, complete variable angle PFGE system with cooling module, includes all gel casting accessories, pump, tubing, DNA standards, 2 plugs of S. cerevisiae DNA size standard, PFGE grade agarose

This product is being discontinued soon. Please contact customer service for more information.

The following replacement is recommended:

  • CHEF Mapper XA Chiller System, 120 V (1703670)
(Discontinued) CHEF-DR II Chiller System

1703725
120 V, complete PFGE system with cooling module, pump, gel casting accessories, 2 plugs S. cerevisiae DNA size standard, 5 g pulsed field Certified agarose, 5 g pulsed field Certified megabase agarose

This product is no longer available. Please contact technical support for more information on replacement product.

The following replacement is recommended:

  • CHEF Mapper XA Chiller System, 120 V (1703670)

Documents

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
6228 Using Pulsed Field Gel Electrophoresis Standards, Rev A Click to download

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