User:Abass Conteh/Notebook/Analysis of CRIPT mutants/Entry Base

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Project name Main project page

Screening Salk T-DNA Mutants

Purpose: Identify plants with T-DNA insert and determine their genotypes.

Extracting Plant DNA

Materials and Reagents Needed:

  DNA extraction materials:
   1. Extraction buffer: 
       200 mM Tris-HCl (pH 8.0)
       200 mM NaCl
       25 mM EDTA
       0.5% SDS.
  2. Sample leaf


 12 µM Gene-specific Forward primer 
 12 µM Gene-specific Reverse primer 
 12 µM LBb1 primer 
 10X Ex-Taq buffer 
 dNTP Mix 
 Ex-Taq DNA polymerase 
 Sterile water 
 1-kb DNA ladder 
 PCR Machine  
 0.2 mL PCR tubes 
 1.5 mL microcentrifuge tubes 
 P-10, P-20, P-200 Pipet 
 PCR rack for 0.2 mL PCR tubes 
 Rack for 1.5 mL microcentrifuge tubes 
 Filtered Pipet tips for PCR 
 Ice bucket 
 Gloves 
 Microcentrifuge 
 Agarose 
 Gel apparatus and power supply 
 Gel Documentation System


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