CLONING 2

title: 'CLONING 2 ' updated: 2023-12-17T12:56:29 created: 2023-12-04T10:40:40

VDAC1 cloning

HindIII digest of plasmids 1,2

• cca 2000 ng each + 1 ul HindIII-HF, 37°C 1h
• 1 ul CIP for plasmid 2
• Gel

1

• 12,5 ul plasmid
• 1 ul HindIII
• 1,5 ul rCutSmart
• 0 ul H2O

2

• 6,9 ul plasmid
• 1 ul Hindiii
• 1,5 ul rcutsmart
• 1 ul cip
• 4,6 ul H2O

1 - 65 ng/ul 80 ng = 1,2 ul 2 - 27 ng/ul 90 ng = 3,33 ul

1 ul T4 1 ul buf 3,45 ul H2O 1:5

Transformation

Mini 6 clones + 1 control

Ecori digest MM = 9 ul rcutsmart 2,25 ul ul ecoriHF

26,2Р h2o

Mixture = 5 ul mm 5 ul dna