restriction enzyme apoi hf  (New England Biolabs)

Journal: bioRxiv

Article Title: Deletion of HindIIR and HindIIIR improves DNA transfer via electroporation to Haemophilus influenzae Rd

doi: 10.1101/2024.07.09.602704

Figure Lengend Snippet: ( A ) Transformation competent cells were prepared from wild-type (“WT”, n = 4) and ΔHindII/III (n = 4) and then electroporated with either pSU20 or pHflu4 plasmid DNA. ΔHindII/III produced more colony forming units (CFU) per microgram of transforming DNA (*: p = 0.12, **: p = 0.04). Error bars represent standard error from the mean. ( B ) Restriction digest of plasmids isolated from ΔHindII/III and WT transformants (+: original plasmid used for transformation, L: NEB 1kb plus ladder)

Article Snippet: Isolated pSU20 and pHflu4 were digested using ApoI-HF (New England Biolabs #R3566) and EcoRI-HF (New England Biolabs # R3101), respectively.

Techniques: Transformation Assay, Plasmid Preparation, Produced, Isolation

Journal: bioRxiv

Article Title: Deletion of HindIIR and HindIIIR improves DNA transfer via electroporation to Haemophilus influenzae Rd

doi: 10.1101/2024.07.09.602704

Figure Lengend Snippet: ( A ) Red fluorescence provided by pHflu4 is visible in ambient light, while green provided by pHflu6 and pHflu7 is better observed under UV light. H. influenzae ΔHindII/III transformants were serially diluted and spot-plated on sBHI supplemented with 5 µg/ml chloramphenicol and grown for 2 days at 37 °C. The image below was taken under UV light. ( B ) H. influenzae ΔHindII/III carrying pHflu1, pHflu4, pHflu6, and pHflu7 as observed by confocal fluorescence microscopy. Red channel excitation/emission wavelength: 594/613 nm; red channel: 488/543 nm

Article Snippet: Isolated pSU20 and pHflu4 were digested using ApoI-HF (New England Biolabs #R3566) and EcoRI-HF (New England Biolabs # R3101), respectively.

Techniques: Fluorescence, Microscopy