Molecular Biology 1st Lab meeting - spring 2002

1) Watch Lab safety video

2) Practice pipetting (see reminder page).

3) Make solutions. What is 1X anyway?

If a graduated cylinder was used for a solution of salts, just rinse it 3 times in distilled water before using. Cylinders that were used with a protein solutions (e.g. media) should be washed with detergent before using again.


Tuesday's Solutions

 sterile yellow and blue tips  
 sterile culture tubes  
 sterile tooth picks  
 LB + amp. agar plates  make 1 L
 5M NaCl  make 100 mL
 1 M Tris (pH 8.0)  make 1 L
 0.5 M EDTA  make 500 mL
 5 M NaOH  make 100 mL
 10X tris saline  make 125 mL
 10X transfer buffer (western)  make 500 mL
 20X SSC  make 1 L
 ECL 2° Wash  make 2 L
 Ampicillin stock  ask Dr. C.
 5X TBE  make 1 L

How much and what kind of container?

 Reagent

 Volume

 Container

 Autoclave?

 sterile yellow tips  1 box per person  yellow box  YES
 sterile culture tubes  1 full blue rack  blue rack  YES
 sterile tooth picks  1 beaker per lab day  small beaker + foil  YES
 LB+amp. agar plates
 1 L
  2 L Flask  YES + amp and pour
 0.5 M EDTA  500 mL  1 L bottle  NO
 5M NaCl  100 mL  100 mL bottle  NO
 5 M NaOH  100 mL  100 mL bottle  NO
 10X tris saline 125 mL  250 mL bottle  NO
 Turbo Denature Buff.  2 L  2 x 1 L bottles  NO
 10X transfer stock (western)  500 mL  500 mL bottle  NO
 20X SSC  1L  1L bottle  NO
 ECL 2° Wash  2 L  2 x 1L bottle  NO
 Ampicillin Stock  ask Dr. C.  blue cap 15 mL tube  filter sterilize
 5X TBE  1 L  1 L bottle  NO


Thursday's Solutions

 sterile yellow tips  
 sterile culture tubes  
 sterile tooth pick  
 LB liq. medium  make 500 mL
 LB + amp agar plates  make 1L
 3 M sodium acetate  make 50 ml you figure this one out (pH to 5.2 with glacial acetic acid)
 Turbo Transfer Buffer  make 2 L
 Turbo Denature Buffer  make 2 L
 Turbo 5X Neutralizing Buffer  make 500 mL
 ECL primary wash (w/o urea)  make 2 L
 ponsceau  make 100 mL
 1X stock of TE  50 mL
 5X TBE  make 1 L

 

How much and what kind of container?

 Reagent

 Volume

 Container

 Autoclave?

 sterile yellow tips  1 box per person  yellow box  YES
 sterile culture tubes  1 full blue rack  blue rack  YES
 sterile tooth picks  1 beaker  small beaker + foil  YES
 LB media, no amp  500 ml  five 100 ml bottles  YES
 LB+amp. agar plates  1L  in a two liter flask  YES + amp and pour
 3 M sodium acetate  50 ml  100 ml bottle  NO pH = 5.2: glacial acetic
 Turbo Denature Buff.  2 L  2 x 1 L bottles  NO
 Turbo Trans. Buf.  2 L  2 X 1 L bottle  NO
 Turbo Neut. Buf.  200 ml  500 ml bottle  NO
 ECL 1° wash  2 L  2 X 1 L bottle  NO
 1X stock of TE  50 mL  100 mL bottle  YES
 ponsceau  100 ml  100 ml bottle  NO
 5X TBE  1 L  1 L bottle  YES


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Send comments, questions, and suggestions to: macampbell@davidson.edu