Dr. David W. Andrews - Lab Manual

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CELL FREE SYSTEMS	> TRANSLATION	> E coli Translation	> Translation Protocol
			> Sample Protocol
			> Troubleshooting
			> Lysate Preparation of S-30
			> Membrane-Free Ribosomes (mfrs)
			> Ribosomal Pellet
			> Dialysis Tubing Work-up
			> Membrane-Free Translation System
			> Buffers and Reagents

Sample Protocol

Get a printable version of this protocol (requires Adobe Acrobat)

Part A:S-30 (with endogenous membranes)

2.5X Reaction buffer	4uL
DNA template	0.5uL
35S Met	1uL
s-30 lysate	3uL
water	1.5uL
Total	10uL

Incubate 45 minutes at 37ºC.

Part B: S-30 (without membranes)

2.5X reaction buffer	4uL
DNA template	0.5uL
35S Met	1uL
S-135 lysate	3uL
membrane free ribosomes	0.1uL
water	1.4uL
Total	10uL

Incubate 45 minutes at 37ºC