Anomalies in the sedimentation of deoxyribonucleic acid from Haemophilus influenzae in alkaline sucrose gradients.
Difficulties were experienced in obtaining reproducible results for the sedimentation in alkaline sucrose gradients of Haemophilus influenzae deoxyribonucleic acid (DNA). The technique of McGrath and Williams of lysing whole cells on top of an alkaline sucrose gradient was employed. The addition of 0.2% sodium lauryl sulfate to the 0.2 n NaOH lysing solution and reduction of the hemin concentration in the growth medium increased the reproducibility to 100% for log-phase cells handled in a manner mimicking the handling procedure used for irradiating cells with ultraviolet light. Distributions of DNA with number average molecular weights of 200 x 10(6) were routinely obtained by using these modifications.
D001433:Bacteriolysis; D002247:Carbon Isotopes; D002499:Centrifugation, Density Gradient / Q000592:standards*; D003470:Culture Media; D004269:DNA, Bacterial / Q000032:analysis*; D004277:DNA, Single-Stranded / Q000032:analysis; D005069:Evaluation Studies as Topic; D006193:Haemophilus influenzae / Q000032:analysis* / Q000254:growth & development; D006418:Heme; D008970:Molecular Weight; D012967:Sodium Dodecyl Sulfate; D012972:Sodium Hydroxide; D013395:Sucrose; D013936:Thymidine; D014316:Tritium
D002247:Carbon Isotopes; D003470:Culture Media; D004269:DNA, Bacterial; D004277:DNA, Single-Stranded; D014316:Tritium; D012967:Sodium Dodecyl Sulfate; D006418:Heme; D012972:Sodium Hydroxide; D013395:Sucrose; D013936:Thymidine