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manual of BAC Protocols in hbz lab (plant genomics)
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Manual of BAC Protocols

Contents

Introduction Construction of Large-Insert Bacterial Clone Libraries
   
Experiment  1   Preparation of High-quality Megabase-size DNA
   
1.1
Preparation of Megabase-size Nuclear DNA from Plants
   
1.2 Preparation of Megabase-size DNA from Animals
   
1.3 Preparation of Megabase-size DNA from Insects
   
Experiment  2 Preparation of BAC and BIBAC Cloning Vectors
   
Experiment  3

Generation of Large DNA Fragments from Megabase DNA Embedded in LMP Agarose by Partial Digestion for BAC and BIBAC Cloing: Determination of Optimal Partial Digestion Conditions

   
Experiment  4 BAC and BIBAC Cloning: Partial Digestion of Megabase DNA, Size Selection, Ligation and Transformation 
   
Experiment  5 BAC Analysis
   
Experiment  6 Large-insert, Ordered Bacterial Clone Library Manipulation
   
Experiment  7 BAC and BIBAC Library Screening
   
Experiment  8 BAC End Isolation
   
Appendix  A Preparation of Yeast Chromosome-size DNA and Lambda DNA Ladder
   
Appendix  B Preparation of Competent Cells for Bacterial Transformation by Electroporation
   
Appendix  C Chemicals and Supplies
   
Appendix  D Solutions and Preparation
   
Appendix  E Restriction Site Maps of Widely Used BAC, BIBAC, PBC and PAC Vectors
   
Appendix  F Nucleotide Sequences:  pBeloBAC11, pCLD04541 and pBACe3.6
   
References Cited  
   
   

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manual of BAC protocols.

 
 
 
 
 
 
 
 
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