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

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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