Check to see if digestion visualization Primers Work

Electrophoresis after restriction enzyme digestion‎

• All 4 of PCR products were purified via Microcon YM-10

Promoter band in lane 6 wasn't visible

• This vector didn't have a primer annealing site for our new primers

Check of Ligation Mixtures

TAKARABIO's Ligation Mixture [I] and Ligation Mixture Mighty Mix [M]

• After PCR and gel extracted pSB1C3 volume of either Ligation Mixture equal to pSB1C3 solution volume was added
• Transformed
• incubated in 200 uL of LB medium
• 100 uL of each solution was plated on mediums containing 34 ug/uL and 12 ug/uL of chloramphenicol

Various Checks

Electrophoresis to check if various DNA manipulations were successful

Band weren't visible in lane 3

• Dimers, trimers and etc were hardly visible maybe severely diluted

In lane 7 had detectable by very weak band

• There weren't any problems with gel extraction