Koeris/Notebook/2007-1-19: Difference between revisions

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=Re-cloning of [His]<sub>6</sub>=
Due to lack of material, I could not recover the A plasmid and have to re-clone it.


==PCR set-up==
*100 ul total reaction volume
*1ul template DNA @ ~40ng/ul
*200pmol primer mix
*97ul Taq SuperMix
===Pipetting scheme===
Labels in the respective field codes
{| border="1"
! Primer !! Tube Label
|-
! A his6 2F&R 200pmol
| A1
|-
! A his6 2F&R 200pmol
| A2
|}
===Thermal profile===
#92 deg C - 5min
#92 deg C - 30s
#50 deg C - 45s
#72 deg C - 120s
#Cycle back to 2. 29X
#72 deg C - 10min
#4 deg C - indefinite
==Gel image==
Loaded 5ul, i.e. 5% of Rx on the gel
==Restriction digest==
*Use Qiagen PCR cleanup kit to remove salts and enzymes
*Digest PCR amplicons with KpnI, Hind III - double digest

Latest revision as of 07:24, 13 February 2007

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