Team:KIT-Kyoto/Notebook/ATF1/may

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ATF1

May 16th

We performed PCR to amplify the ATF1 gene.

Primers:

Forward:5’-TACGCCTCGAGATGAATGAAATCGATGAGAAAAATCAGGCCCC-3’

Reverse:5’-ATATTGCTTAGCAGGGCCTAAAAGGAGAGCTTTGTAAATGGAGCAAAGC-3’

Reaction composition is as follows:

Buffer

50µL

dNTP

20µL

Primer mix

1µL

DNA sample

0.5µL

KOD-FX

2µL

H2O

26.5µL

total

100µL

We could not get any PCR product.

 

 

May 17th -20th

Changed PCR conditions with another DNA template. We performed PCR of the ATF1 gene again.

Primers:

Forward :5’-TACGCCTCGAGATGAATGAAATCGATGAGAAAAATCAGGCCCC-3’

Reverse:5’-ATATTGCTTAGCAGGGCCTAAAAGGAGAGCTTTGTAAATGGAGCAAAGC-3’

Reaction composition is as follows:

Buffer

50µL

dNTP

20µL

Primer mix

1µL

DNA sample

0.5µL

KOD-FX

2µL

H2O

26.5µL

total

100µL

We could not get any PCR product.

 

 

 

May 30th -31th

We transformed a plasmid carrying the ATF1 gene into E.coli.

This plasmid was obtained from iGEM DNA distributions kit 2012 (plate 2, o-7).

We could see approximately 50 colonies and cultured furthermore.