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

50uL

dNTP

20uL

Primer mix

1uL

DNA sample

0.5uL

KOD-FX

2uL

H2O

26.5uL

total

100uL

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

50uL

dNTP

20uL

Primer mix

1uL

DNA sample

0.5uL

KOD-FX

2uL

H2O

26.5uL

total

100uL

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.