Lab 208

Main purpose

PCR for HAO with USER endings after purification

Who was in the lab

Henrike, Ariadni

Procedure

Gel purification

of HAO User (faint bands) and HAO ex (bright bands). HAO ex will be used as a template for new PCRs to amplify HAO User.

PCR for HAO with USER endings

Template: HAO extraction fragment (purified from PCR run on 04.09)

Primers: 18a, 18b

Two versions, one with 5% DMSO and one with 3% DMSO and two programs: Touchdown (62C -> 55) and a single annealing temperature of 59C.

PCR reaction master mix (x5)

• Add 49 ul of the master mix, and
• Add 1 ul DMSO (for the 5%) or 1 ul of MQ (for the 3%)

Results

Gel on yesterdays PCR

• 2-log ladder
• HAO USER on 59C, 3% DMSO
• HAO USER on 59C, 3% DMSO
• HAO USER on 57C, 3% DMSO
• HAO USER on 57C, 3% DMSO
• HAO USER on touchdown
• HAO USER on touchdown
• HAO USER on touchdown, 3% DMSO
• HAO USER on touchdown, 3% DMSO
• HAO extraction fragment on 57C, template -20 culture
• HAO extraction fragment on 57C, template -20 culture
• HAO extraction fragment on 57C, template -80 culture
• HAO extraction fragment on 57C, template -80 culture
• HAO extraction fragment on 59C, template -20 culture
• HAO extraction fragment on 59C, template -20 culture
• HAO extraction fragment on 59C, template -80 culture
• HAO extraction fragment on 59C, template -80 culture
• 2-log ladder

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