Material and Equipment

DNA EXTRACTION:

 Cheek Cell lysis Buffer (50mM TrisHCL, SDS 1%)

 Proteinase K
 5M NaCL
 Saline solution
 100% and 70% cold ethanol
 Distilled water
 200-1,000 ul micropipettes and tips.
 Disposable transfer pipets
 1.5ml ependorf tubes.
 Water bath at 50 degrees C
 Centrifuge-15ml Falcon tube.
 Cups to spit in.

AGAROSE GEL ELECTROPHORESIS OF DNA:

 PCR products from Lab2

 SB buffer (0.01M NaOH and 0.36M Boric acid, pH 8)
 10mg/ml ethidium bromide 10 000X Gelred
 1kp DNA ladder
 Gel loading buffer (60% glycerol, 0.2% Orange sodium)
 Agarose
 Distilled water
 250 ml bottle (agarose gel used only)
 Microwaves
 UV lamps & camera
 10 µl micropipette
 Agarose gel electrophoresis apparatus

POLYMERASE CHAIN REACTION:

 DNA from cheek cell

 Primer HGH I
 Primer HGH II
 Toptaq master mix
 Distilled water
 PCR machine
 1.5 ml eppendorf tube
 PCR tube 0.2ml
 2 µl, 100 µl, & 1000 µl micropipette
PCR PURIFICATION:

 Wizard SV Gel
 PCR Clean-up System (Promega)

AGAROSE GEL ELECTROPHORESIS OF PCR PURIFICATION:

 PCR purification products

 SB buffer (0.01M NaOH and 0.36M Boric acid, pH 8)
 10mg/ml ethidium bromide 10 000X Gelred
 100bp DNA ladder
 Gel loading buffer (60% glycerol, 0.2% Orange sodium)
 Agarose
 Distilled water
 250 ml bottle (agarose gel used only)
 Microwaves
 UV lamps & camera
 10 µl micropipette
 Agarose gel electrophoresis apparatus

DNA CLONING – DNA RECOMBINATION:

 PCR product from Lab 4

 2X ligation buffer
 pGEM-T-Easy vector
 T4 DNA ligase
 1.5 ml eppendorf tube
 2 µl & 10 µl micropipette

DNA CLONING – TRANSFORMATION:

 pGEMT-easy vector system II

 LB with ampicillin
 LB without ampicillin
 S.O.C medium
 Water bath (420 C)
 Ice box

DNA CLONING – PLASMID PURIFICATION:

 QIAprep Miniprep Kits

 LB medium
 Antibiotic, e.g Ampicillin
 70% ethanol
AGAROSE GEL ELECTROPHORESIS OF PLASMID PURIFICATION:

 Plasmid DNA
 SB buffer (0.01M NaOH and 0.36M Boric acid, pH 8)
 10mg/ml ethidium bromide 10 000X Gelred
 100bp DNA ladder
 Gel loading buffer (60% glycerol, 0.2% Orange sodium)
 Agarose
 Distilled water
 250 ml bottle (agarose gel used only)
 Microwaves
 UV lamps & camera
 10 µl micropipette
 Agarose gel electrophoresis apparatus