pBAR_PgpdA transformed into competent cell

pBAR_PgpdA containing cultures used in 2 in 1

pBAR_PgpdA extracted and checked with plasmid PCR and restriction digestion

pBAR_PgpdA plasmid stored and transformed cultures cryopreserved

pBAR_PgpdA digested for ligation with pVP-EL222 and TtrpC

pBAR_PgpdA_EL222_TtrpC transformed into competent cell

pBAR_PgpdA_EL222_TtrpC extracted and checked with restriction digestion (failure)

pBAR_PgpdA_EL222_TtrpC transformed into competent cell

pBAR_PgpdA_EL222_TtrpC containing cultures used in 3 in 1

pBAR_PgpdA_EL222_TtrpC checked with restriction digestion

pBAR_PgpdA_EL222_TtrpC amplified with plasmid PCR

EL222 and TtrpC ran ligation PCR ------- NC had band

EL222 and TtrpC ran ligation PCR ------ with remixed primer

EL222 and TtrpC ran ligation PCR ------ the concentration was not high enough after gel extraction

EL222 and TtrpC ran ligation PCR

EL222 and TtrpC ran ligation PCR

EL222-TtrpC checked with restriction digestion

pBAR_PgpdA digested for ligation with EL222_TtrpC

pBAR_PgpdA digested for ligation with EL222_Ttrp

pBAR_PgpdA_EL222_TtrpC transformed into competent cell

pBAR_PgpdA_EL222_TtrpC containing cultures used in 3 in 1

pBAR_PgpdA_EL222_TtrpC extracted from transformed cultures

pBAR_PgpdA_EL222_TtrpC amplified with plasmid PCR

pBAR_PgpdA_EL222_TtrpC checked with restriction digestion (failure)

pBAR_PgpdA_EL222_TtrpC checked with restriction digestion (we changed enzymes)

EL222-TtrpC digested with restriction enzyme

pBAR_PgpdA_EL222_TtrpC checked with restriction digestion

pBAR_PgpdA_EL222_TtrpC transformed into competent cell( there was no E.coli on our first plate )

pBAR_PgpdA_EL222_TtrpC transformed into competent cell

pBAR_PgpdA_EL222_TtrpC containing culture used in 3 in 1

pBAR_PgpdA_EL222_TtrpC checked with restriction digestion

pBAR_PgpdA_EL222_TtrpC amplified with plasmid PCR

Successfully constructed the plasmid pBAR_PgpdA_EL222_TtrpC

pBAR_PgpdA_EL222_TtrpC digested for transforming into Metarhizium anisopliae