Difference between revisions of "Bryan Hernandez/20.109/Lab notebook/Module 1/Day 3"
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==Ligations and Transformations== | ==Ligations and Transformations== | ||
+ | --[http://openwetware.org/wiki/User:Bryanh |Bryanh] 17:17, 16 February 2007 (EST) | ||
==Purpose== | ==Purpose== | ||
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==Protocols== | ==Protocols== | ||
When you have a break from the work described below, be sure to examine the plaques you plated last time. Record the number of plaques on each plate, their appearance and any observations or conclusions you can draw. | When you have a break from the work described below, be sure to examine the plaques you plated last time. Record the number of plaques on each plate, their appearance and any observations or conclusions you can draw. | ||
+ | |||
+ | ===Part 0=== | ||
+ | '''counting PFUs:''' | ||
+ | |||
+ | ==Phage infection== | ||
+ | {| border="1" cellpadding="2" | ||
+ | !width="10"|A | ||
+ | !width="10"|B | ||
+ | !width="10"|C | ||
+ | |- | ||
+ | |'''Phage dilution'''||'''Phage Strain'''||'''PFUs Counted'''|| | ||
+ | |- | ||
+ | |none||E4||lawn|| | ||
+ | |- | ||
+ | |none||K07||lawn|| | ||
+ | |- | ||
+ | |10^-4||E4||too many to count|| | ||
+ | |- | ||
+ | |10^-4||K07||too many to count|| | ||
+ | |- | ||
+ | |10^-6||E4||2000|| | ||
+ | |- | ||
+ | |10^-6||K07||no PFUs (this is probably an error in our dilution step)|| | ||
+ | |- | ||
+ | |} | ||
+ | |||
+ | |||
+ | |||
===Part 1: Ligation reactions=== | ===Part 1: Ligation reactions=== | ||
− | For your ligation, you will mix the M13K07 backbone you prepared with the annealed oligonucleotides. As control reactions you will also prepare a | + | For your ligation, you will mix the M13K07 backbone you prepared with the annealed oligonucleotides. As control reactions you will also prepare a |