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微信小章| 产品名称： | lambda EMBL12 |
|---|---|
| 商品货号： | TS195092 |
| Designations： | lambda EMBL12 |
| Depositors： | G Scherer |
| Biosafety Level： | 1 |
| Vector Information： | Size (kb): 43.2000007629394500 Vector: lambda EMBL12 (phage, lambda - replacement) Construction: lambda EMBL3 pUC12 Construct size (kb): 43.20000076293945 Features: insert detection: Spi+ replicon: lambda |
| Applications： | vector for constructing genomic libraries vector permitting positive selection for inserts |
| Comments： | Phage with inserts have the Spi- phenotype. A bacteriophage lambda vector for construction and screening of genomic libraries with a cloning capacity of 8-23 kb. |
| References： | Natt E, Scherer G. EMBL12, a new lambda replacement vector with sites for SalI, XbaI, BamHI, SstI and EcoRI. Nucleic Acids Res. 14: 7128, 1986. PubMed: 3020507 |
| Shipped： | freeze-dried |
| Shipping Information： | Shipped: xa0Freeze dried bacteria-free phage lysate. Titering: 1.xa0xa0xa0xa0xa0xa0xa0xa0xa0 Make fresh plating bacteria.xa0 Grow E. coli host strain overnight or at least to A600 = 0.4 in medium containing 0.2% maltose (to give higher titers). 2.xa0xa0xa0xa0xa0xa0xa0xa0xa0 Spin down cells in a low speed centrifuge.xa0 Resuspend in 0.4 volumes 10 mM MgSO4 or SM buffer.xa0 Store at 40C. 3.xa0xa0xa0xa0xa0xa0xa0xa0xa0 Dilute phage in 10 mM MgSO4 xa0of SM buffer. Mix gently because vigorous mixing reduces the titer. 4.xa0xa0xa0xa0xa0xa0xa0xa0xa0 Add 100 ul phage dilution to 100 ul prepared plating bacteria and mix gently.xa0 Incubate in a 370C water bath for 20 minutes to allow phage to adsorb. 5.xa0xa0xa0xa0xa0xa0xa0xa0xa0 Add 3 ml LB lambda top agar (see below) containing 0.2% maltose and mix gently.xa0 Pour onto plates.xa0 Incubate overnight at 370C.xa0 Fresh plates give larger plaques. Storage: Libraries can be frozen in liquid nitrogen with no significant loss in titer, even after repeated freeze-thaw cycles.xa0 The libraries on dry ice can be stored at 40C or can be kept frozen at -70 0C or in liquid nitrogen.xa0 Always freeze by plunging into liquid nitrogen. LB Lambda top agar medium: NaClxa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0 5 g Tryptonexa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0 xa010 g Yeast extractxa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0 5 g Distilled water toxa0xa0xa0xa0xa0xa0xa0 1 L Sterilize at 1210C, 15 minutes.xa0 Cool to approximately 500C and add the following sterile solutions. 1M CaCl2xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0 5 ml MgSO4 H2Oxa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0 to a final concentration of 0.2% w/v 50% maltosexa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0xa0 5 ml For solidxa0 media, add 7 g. agar or agarose (for top agar) per liter or 15 g agar for basal medium prior to autoclaving. Reference: Biotechniques 5: 724-728, 1987. xa0xa0xa0xa0 |