| 1、The BAC clone from glycerol culture sublibrary is innoculated into 3 ml of LB with 20µg/ml chloramphenicol and cultured at 37°C with shaking for 16hrs. The liquid bacterial culture is transferred into the tube specific for AutoGene 740 DNA Isolation System (Autogen, Inc. ) |
Step No. | Process | Parameter set |
---|---|---|
07 | Agitation to resuspend cell pellet | 80 Hz; 5 minutes |
19 | Agitation to precipitate DNA | 90 Hz; 5 minutes |
25 | Centrifuge to pellet DNA | 3500 rpm; 2 minutes |
33 | Agitation to dissolve DNA | 1Hz; 1 amp; 1 Second |
5、Common yield of this approach is about 2µg BAC DNA / 3ml Bacterial culture, but the quantity and quality of DNA are dependended on the BAC clone itself such as insert size etc. You can test DNA quality and quantity by digesting some of it with Hind III and running a 1% agarose gel for 2-3 hrs.
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