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Determination of the minimal functional region of CEN 4. ( A ) Transformation results for subclones that resulted from NotI digestion after PCR with primers containing a NotI site , followed by religation and transformation of purified plasmids from E. coli plasmid preparations. Transformation with plasmid subclones containing increasingly smaller CEN 4 regions from p1120 revealed that while large colonies (which are integrative transformants) remain the same size, after NotI subclone 6188, small colonies become increasingly smaller as the CEN 4 region is reduced. Subclone 3684 small colonies are barely distinguishable from p1114, which contains the NAT marker only. Bar = 3 mm. ( B ) Transformation results for subclones that resulted from SacII digestion, religation, and transformation, as above. The SacII subclones were derived from the opposite site of CEN 4 and showed small colony size decreasing after the 2963 subclone. Plates were photographed at 2.5×, with an exposure time of 110 ms. Bar = 3 mm.
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escherichia coli one shot top 10 electrocompetent cells - by Bioz Stars, 2026-09
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top 10 escherichia coli cells  (Thermo Fisher)
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Determination of the minimal functional region of CEN 4. ( A ) Transformation results for subclones that resulted from NotI digestion after PCR with primers containing a NotI site , followed by religation and transformation of purified plasmids from E. coli plasmid preparations. Transformation with plasmid subclones containing increasingly smaller CEN 4 regions from p1120 revealed that while large colonies (which are integrative transformants) remain the same size, after NotI subclone 6188, small colonies become increasingly smaller as the CEN 4 region is reduced. Subclone 3684 small colonies are barely distinguishable from p1114, which contains the NAT marker only. Bar = 3 mm. ( B ) Transformation results for subclones that resulted from SacII digestion, religation, and transformation, as above. The SacII subclones were derived from the opposite site of CEN 4 and showed small colony size decreasing after the 2963 subclone. Plates were photographed at 2.5×, with an exposure time of 110 ms. Bar = 3 mm.
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Determination of the minimal functional region of CEN 4. ( A ) Transformation results for subclones that resulted from NotI digestion after PCR with primers containing a NotI site , followed by religation and transformation of purified plasmids from E. coli plasmid preparations. Transformation with plasmid subclones containing increasingly smaller CEN 4 regions from p1120 revealed that while large colonies (which are integrative transformants) remain the same size, after NotI subclone 6188, small colonies become increasingly smaller as the CEN 4 region is reduced. Subclone 3684 small colonies are barely distinguishable from p1114, which contains the NAT marker only. Bar = 3 mm. ( B ) Transformation results for subclones that resulted from SacII digestion, religation, and transformation, as above. The SacII subclones were derived from the opposite site of CEN 4 and showed small colony size decreasing after the 2963 subclone. Plates were photographed at 2.5×, with an exposure time of 110 ms. Bar = 3 mm.
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Determination of the minimal functional region of CEN 4. ( A ) Transformation results for subclones that resulted from NotI digestion after PCR with primers containing a NotI site , followed by religation and transformation of purified plasmids from E. coli plasmid preparations. Transformation with plasmid subclones containing increasingly smaller CEN 4 regions from p1120 revealed that while large colonies (which are integrative transformants) remain the same size, after NotI subclone 6188, small colonies become increasingly smaller as the CEN 4 region is reduced. Subclone 3684 small colonies are barely distinguishable from p1114, which contains the NAT marker only. Bar = 3 mm. ( B ) Transformation results for subclones that resulted from SacII digestion, religation, and transformation, as above. The SacII subclones were derived from the opposite site of CEN 4 and showed small colony size decreasing after the 2963 subclone. Plates were photographed at 2.5×, with an exposure time of 110 ms. Bar = 3 mm.
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Determination of the minimal functional region of CEN 4. ( A ) Transformation results for subclones that resulted from NotI digestion after PCR with primers containing a NotI site , followed by religation and transformation of purified plasmids from E. coli plasmid preparations. Transformation with plasmid subclones containing increasingly smaller CEN 4 regions from p1120 revealed that while large colonies (which are integrative transformants) remain the same size, after NotI subclone 6188, small colonies become increasingly smaller as the CEN 4 region is reduced. Subclone 3684 small colonies are barely distinguishable from p1114, which contains the NAT marker only. Bar = 3 mm. ( B ) Transformation results for subclones that resulted from SacII digestion, religation, and transformation, as above. The SacII subclones were derived from the opposite site of CEN 4 and showed small colony size decreasing after the 2963 subclone. Plates were photographed at 2.5×, with an exposure time of 110 ms. Bar = 3 mm.
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tiangen biotech co escherichia coli top 10 competent cell
Determination of the minimal functional region of CEN 4. ( A ) Transformation results for subclones that resulted from NotI digestion after PCR with primers containing a NotI site , followed by religation and transformation of purified plasmids from E. coli plasmid preparations. Transformation with plasmid subclones containing increasingly smaller CEN 4 regions from p1120 revealed that while large colonies (which are integrative transformants) remain the same size, after NotI subclone 6188, small colonies become increasingly smaller as the CEN 4 region is reduced. Subclone 3684 small colonies are barely distinguishable from p1114, which contains the NAT marker only. Bar = 3 mm. ( B ) Transformation results for subclones that resulted from SacII digestion, religation, and transformation, as above. The SacII subclones were derived from the opposite site of CEN 4 and showed small colony size decreasing after the 2963 subclone. Plates were photographed at 2.5×, with an exposure time of 110 ms. Bar = 3 mm.
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Determination of the minimal functional region of CEN 4. ( A ) Transformation results for subclones that resulted from NotI digestion after PCR with primers containing a NotI site , followed by religation and transformation of purified plasmids from E. coli plasmid preparations. Transformation with plasmid subclones containing increasingly smaller CEN 4 regions from p1120 revealed that while large colonies (which are integrative transformants) remain the same size, after NotI subclone 6188, small colonies become increasingly smaller as the CEN 4 region is reduced. Subclone 3684 small colonies are barely distinguishable from p1114, which contains the NAT marker only. Bar = 3 mm. ( B ) Transformation results for subclones that resulted from SacII digestion, religation, and transformation, as above. The SacII subclones were derived from the opposite site of CEN 4 and showed small colony size decreasing after the 2963 subclone. Plates were photographed at 2.5×, with an exposure time of 110 ms. Bar = 3 mm.

Journal: Journal of Fungi

Article Title: Development of a Shuttle Vector That Transforms at High Frequency for the Emerging Human Fungal Pathogen: Candida auris

doi: 10.3390/jof10070477

Figure Lengend Snippet: Determination of the minimal functional region of CEN 4. ( A ) Transformation results for subclones that resulted from NotI digestion after PCR with primers containing a NotI site , followed by religation and transformation of purified plasmids from E. coli plasmid preparations. Transformation with plasmid subclones containing increasingly smaller CEN 4 regions from p1120 revealed that while large colonies (which are integrative transformants) remain the same size, after NotI subclone 6188, small colonies become increasingly smaller as the CEN 4 region is reduced. Subclone 3684 small colonies are barely distinguishable from p1114, which contains the NAT marker only. Bar = 3 mm. ( B ) Transformation results for subclones that resulted from SacII digestion, religation, and transformation, as above. The SacII subclones were derived from the opposite site of CEN 4 and showed small colony size decreasing after the 2963 subclone. Plates were photographed at 2.5×, with an exposure time of 110 ms. Bar = 3 mm.

Article Snippet: Escherichia coli One Shot TOP 10 Electrocompetent cells (Invitrogen, Carlsbad, CA, USA) were utilized for all bacterial transformations.

Techniques: Functional Assay, Transformation Assay, Purification, Plasmid Preparation, Marker, Derivative Assay

Stability of p1153. ( A ) Transformation of E. coli with total DNA prepared from transformed large (L)- and small (S)-colony DNA of C. auris, showing that only small-colony total DNA contains episomal plasmids, which are capable of being rescued by transformation of E. coli . ( B ) EcoRI restriction digest pattern of p1153 (6.6 kb) rescued from E. coli after DNA extraction from small-colony C. auris transformants. L = ladder, C = control p1153 plasmid, 1–4 plasmids rescued from E. coli after transformation with small-colony C. auris total DNA. All plasmids show identical bands of 3.4 kb, 2.2 kb, 0.59 kb, and 0.42 kb (11 bp band is not visible). ( C ) XbaI digestion of p1210 (11.6 Kb) showing predicted bands of 6.0 kb, 3.9 Kb, and 1.7 Kb. L = ladder, C = control p1210 plasmid, 1–10 plasmids rescued in E. coli after transformation with small-colony C. auris total DNA. ( D ) XbaI digestion of p1213 (16.6 Kb) showing predicted bands of 7.0 kb, 5.7 Kb, and 3.9 Kb. L = ladder, C = control p1210 plasmid, 1–10 plasmids rescued from E. coli after transformation with small-colony C. auris total DNA. ( E ) Transformation frequencies of p1153, p1210, and p1213, which corresponded to size. p1153 is the smallest plasmid, while p1213 is the largest. The results suggest that plasmid size influences transformation frequency, which would be expected; however, although p1153 yielded significantly more transformants than the two larger plasmids ( p < 0.05 by ANOVA), the transformation frequency was still good for all three plasmids. Significance level was **** = p < 0.0001.

Journal: Journal of Fungi

Article Title: Development of a Shuttle Vector That Transforms at High Frequency for the Emerging Human Fungal Pathogen: Candida auris

doi: 10.3390/jof10070477

Figure Lengend Snippet: Stability of p1153. ( A ) Transformation of E. coli with total DNA prepared from transformed large (L)- and small (S)-colony DNA of C. auris, showing that only small-colony total DNA contains episomal plasmids, which are capable of being rescued by transformation of E. coli . ( B ) EcoRI restriction digest pattern of p1153 (6.6 kb) rescued from E. coli after DNA extraction from small-colony C. auris transformants. L = ladder, C = control p1153 plasmid, 1–4 plasmids rescued from E. coli after transformation with small-colony C. auris total DNA. All plasmids show identical bands of 3.4 kb, 2.2 kb, 0.59 kb, and 0.42 kb (11 bp band is not visible). ( C ) XbaI digestion of p1210 (11.6 Kb) showing predicted bands of 6.0 kb, 3.9 Kb, and 1.7 Kb. L = ladder, C = control p1210 plasmid, 1–10 plasmids rescued in E. coli after transformation with small-colony C. auris total DNA. ( D ) XbaI digestion of p1213 (16.6 Kb) showing predicted bands of 7.0 kb, 5.7 Kb, and 3.9 Kb. L = ladder, C = control p1210 plasmid, 1–10 plasmids rescued from E. coli after transformation with small-colony C. auris total DNA. ( E ) Transformation frequencies of p1153, p1210, and p1213, which corresponded to size. p1153 is the smallest plasmid, while p1213 is the largest. The results suggest that plasmid size influences transformation frequency, which would be expected; however, although p1153 yielded significantly more transformants than the two larger plasmids ( p < 0.05 by ANOVA), the transformation frequency was still good for all three plasmids. Significance level was **** = p < 0.0001.

Article Snippet: Escherichia coli One Shot TOP 10 Electrocompetent cells (Invitrogen, Carlsbad, CA, USA) were utilized for all bacterial transformations.

Techniques: Transformation Assay, DNA Extraction, Control, Plasmid Preparation