Materials and Reagents

1. The prokaryotic expression vectors: pGEX-6P-1 (GE Healthcare) or pTXB3 (New England Biolabs)
   
2. The expression strain [Escherichia coli (E.coli) BL21 (DE3) or others]
   
3. T-Vector pGEMT-easy and T4-DNA ligase (Promega Corporation, catalog number: A1360 )
   
4. DNA polymerase (Takara Bio Company, catalog number: DR001 )
   
5. Sterile ddH₂O (double distilled water)
   
6. Oligonucleotides (Pitzschke et al., 2009; Wang et al., 2014)
   
7. Chitin resin (New England Biolabs, catalog number: S6651S )
   
8. PBS buffer (see Recipes)
   
9. Column buffer (see Recipes)
   
10. RDSA buffer (Pitzschke et al., 2009) (see Recipes)
    
    Table 1. Oligonucleotides for RDSA
    

    Oligonucleotides for RDSA
    RDSA_1	TAGTTGAGTCTCACAAACGAACAC(N20)CATTCCAAAATCCATGGCTGATA
    RDSA_1 fo	TAGTTGAGTCTCACAAACGAACAC
    RDSA_1 re	TATCAGCCATGGATTTTGGAATG
    RDSA_2	AATGGATCCAAGCTTAAGC(N18)CGTTGAATTCCCATGGACA
    RDSA_2 fo	AATGGATCCAAGCTTAAGC
    RDSA_2 re	TGTCCATGGGAATTCAACG

Equipment

1. Glutathione sepharose beads (GE Healthcare)
   
2. The empty chromatography columns (Bio-Rad Laboratories)
   
3. 4 °C refrigerator
   
4. Centrifuge
   
5. Whirling shaker
   

Procedure