人妻少妇偷人精品无码丨色婷婷av久久久久久久丨欧美xxxx做受性欧美88丨欧洲女人牲交视频免费丨亚洲精品久久久av无码专区

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 bacteria-free phage lysate.

Titering:

1.          Make fresh plating bacteria.? Grow E. coli host strain overnight or at least to A₆₀₀ = 0.4 in medium containing 0.2% maltose (to give higher titers).

2.          Spin down cells in a low speed centrifuge.? Resuspend in 0.4 volumes 10 mM MgSO₄ or SM buffer.? Store at 4⁰C.

3.          Dilute phage in 10 mM MgSO₄ ?of SM buffer. Mix gently because vigorous mixing reduces the titer.

4.          Add 100 ul phage dilution to 100 ul prepared plating bacteria and mix gently.? Incubate in a 37⁰C water bath for 20 minutes to allow phage to adsorb.

5.          Add 3 ml LB lambda top agar (see below) containing 0.2% maltose and mix gently.? Pour onto plates.? Incubate overnight at 37⁰C.? 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.? The libraries on dry ice can be stored at 4⁰C or can be kept frozen at -70 ⁰C or in liquid nitrogen.? Always freeze by plunging into liquid nitrogen.

LB Lambda top agar medium:

NaCl????????????????????????? 5 g

Tryptone???????????????? ?10 g

Yeast extract???????????? 5 g

Distilled water to??????? 1 L

Sterilize at 121⁰C, 15 minutes.? Cool to approximately 50⁰C and add the following sterile solutions.

1M CaCl₂?????????????????? 5 ml

MgSO₄ H₂O??????????????? to a final concentration of 0.2% w/v

50% maltose????????????? 5 ml

For solid? 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.

????