Yu:homemade taq: Difference between revisions
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(PCR recipe added) |
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{| border="1" cellpadding="10" cellspacing="0" | {| border="1" cellpadding="10" cellspacing="0" | ||
|width="200px" valign="top"|'''Reagent''' | |width="200px" valign="top"|'''Reagent''' | ||
|width=" | |width="75px" valign="top"|'''1.5x''' | ||
|width=" | |width="75px" valign="top"|'''2.5x''' | ||
|width=" | |width="75px" valign="top"|'''3.5x''' | ||
|width=" | |width="75px" valign="top"|'''6.5x''' | ||
|width=" | |width="75px" valign="top"|'''9.5x''' | ||
|width=" | |width="75px" valign="top"|'''12.5x''' | ||
|- | |- | ||
|Fwd. Primer 10 μM | |Fwd. Primer 10 μM |
Latest revision as of 12:49, 13 September 2009
Colony PCR
Reagent | 1.5x | 2.5x | 3.5x | 6.5x | 9.5x | 12.5x |
Fwd. Primer 10 μM | 1.5 μL | 2.5 μL | 3.5 μL | 6.5 μL | 9.5 μL | 12.5 μL |
Rev. Primer 10 μM | 1.5 μL | 2.5 μL | 3.5 μL | 6.5 μL | 9.5 μL | 12.5 μL |
10 mM ea dNTPs | 1.5 μL | 2.5 μL | 3.5 μL | 6.5 μL | 9.5 μL | 12.5 μL |
DNA Template | 5 μL of Lyse'n Go per reaction; Lyse'n Go thermocycle takes ~18 min. Can start Lyse'n Go cycle before making master mix | |||||
10x Rxn Buffer (Invitrogen (-MgCl2)) | 7.5 μL | 12.5 μL | 17.75 μL | 32.5 μL | 47.5 μL | 62.5 μL |
50 mM MgCl2 | 2.25 μL | 3.75 μL | 5.25 μL | 9.75 μL | 14.25 μL | 18.75 μL |
milliQ H2O | 45.75 μL | 76.25 μL | 106.75 μL | 198.25 μL | 289.75 μL | 381.25 μL |
Homemade Taq | 7.5 μL | 12.5 μL | 17.5 μL | 32.5 μL | 47.5 μL | 62.5 μL |
Add 45 μL of reaction master mix to each Lyse'n Go reaction |