lab 208

Main purpose

Who was in the lab

Kristian, Julia, Henrike

Procedure

PCR for AMO with USER endings

primers: 17a, 17b

template: gel purified AMO extraction fragment

Program: Standard with 54C annealing temperature and 3:00 extension time

Reaction Mix

PCR for extraction of Nir1

primers: 41a, 41b

template: colony from plate. One colony was dissolved in 100 uL of MilliQ and 1 uL of this solution was taken as template

program: Touchdown PCR analog to program used on 01-08-2013 to amplify Nir.

Reaction Mix

Comment

Both PCRs mentioned above were successful when DMSO was added.

Results

Gel on yesterdays PCR

• 1kb ladder
• Yesterdays sample 1
• Yesterdays sample 2
• Yesterdays sample 3
• Yesterdays sample 4
• Neg.
• Neg.
• Nir2
• cycAX Histag
• 1kb ladder

Gel on ON PCR samples

• 1kb ladder
• Nir1
• Nir1 5%DMSO
• Nir1 1M Betaine
• Nir1 3M Betaine
• Nir2 3M Betaine U
• Nir2 1M Betaine U
• Nir2 5%DMSO U
• Nir2 U
• Neg (Nir)
• Ref 0%DMSO
• Ref 2%DMSO
• Ref 5%DMSO
• AMO U
• AMO 2%DMSO U
• Neg (AMO)
• NirG 5%DMSO
• NirG
• 1kb ladder

Gel on todays PCR samples

• 1kb ladder
• Nir Q5 poly
• Nir 2%DMSO Q5 poly
• Nir 3M Betaine Q5 poly
• Nir 5%DMSO Q5 poly
• Nir1 Q5 poly
• Nir1 2%DMSO Q5 poly
• Nir1 5%DMSO Q5 poly
• Nir1 3M Betaine Q5 poly
• AMO USER primers
• AMO +DMSO USER primers
• AMO +Betaine USER primers
• Nir1 DMSO
• Nir1 Betaine
• 1kb ladder

Conclusion

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