SEED/2010/Day 2

Morning
The purpose of this exercise is to learn to run a restriction digest and an agarose gel to obtain information about a DNA construct.

For each of the two DNA samples, you will set up 4 restriction digests using combinations of the enzymes EcoRI and PstI:
 * 20ul reaction:
 * 2ul NEB2
 * 0.2ul BSA
 * 2ul DNA (~500ng)
 * 16ul DI water
 * 0.5ul per enzyme
 * 4 reactions
 * no enzyme
 * EcoRI only
 * PstI only
 * EcoRI and PstI
 * Incubate at 37C over lunch

Prepare and pour a 1% agarose gel with 10-well combs.

Afternoon

 * Run a gel
 * Cover gel with buffer
 * Remove gel combs
 * Add gel loading buffer to your restriction digests
 * Carefully load samples
 * Load the DNA ladder
 * look at plates from ../Day 1/ and take [[Media:SEED_Quiz1_Antibiotics_MixedCultures.doc‎|quiz]]
 * Image the gel and analyze bands

Lecture

 * BioBricks & Registry of Standard Biological Parts
 * DNA / restriction enzymes
 * iGEM eau d'e colidemo, UT Photography
 * Central Dogma Pardigm
 * DNA->RNA->protein (an enzyme that makes scent)
 * promoters (transcription), RBS (translation)

Instructor Preparation

 * Prepare minipreps of several different BioBrick constructs in the same plasmid but with noticeably different insert lengths
 * Grow up batch culture of registry:Part:BBa_J45200 with 10mM isoamyl alcohol