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Table 1 Number of transformants obtained on indicated selection media

From: Genomic deletions in Aureobasidium pullulans by an AMA1 plasmid for gRNA and CRISPR/Cas9 expression

Transformation1

Plasmid2 (µg)

V proto3 (mL)

Hygromycin B colonies4 (CFU in colonies/µg DNA)5

5-FOA8

Fresh6

Frozen7

1

2

5.2

NT

150 (75)

105 out of 126 (83%)

1

5

5.2

NT

250 (50)

2

2

0.2

0

0

33 out of 33 (100%)

2

2

0.5

10 (52)

5 (26)

2

2

4.5

290 (168)

37 (21)

2

5

0.2

0

0

2

5

0.5

47 (98)

7 (15)

2

5

4.5

572 (132)

116 (27)

  1. NT Not tested
  2. 1Number of the transformation
  3. 2Quantity of plasmid DNA added during the transformation
  4. 3Volume of the used protoplast solution plated with agar (the total volume of the solution is 5.2 mL)
  5. 4Number of colonies on the hygromycin B selection plate and
  6. 5CFU in colonies per µg DNA given in brackets and calculated based on Eq. 1
  7. 6The protoplasts used for the transformation were freshly prepared
  8. 7The protoplasts used were thawed
  9. 8Number of colonies on the 5-FOA selection plate after transfer from the hygromycin B plate. The results are expressed in number of colonies growing out of the total number of colonies transferred, and the representative percentage is also given
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Fungal Biology and Biotechnology

ISSN: 2054-3085

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