Haynes Lab:Notebook/Engineering PC-TFs/2013/03/27: Difference between revisions
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|style="background-color: # | |style="background-color: #FFCC00"|<br>[[Image:Hayneslab3.gif|200px|center]][[Image:Asu logo 3.gif|200px|center]]<span style="font-size:22px;"><br></span> | ||
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==3/27/2013== | ==3/27/2013== | ||
*1. Kozak,V0120 + BL01 | *1. Kozak,V0120 + BL01 | ||
*2. Kozak,V0120 + BL05 | *2. Kozak,V0120 + BL05 | ||
*3. Kozak,V0120 + BL09 | *3. Kozak,V0120 + BL09 | ||
*4. Kozak,V0120 + BL12 | *4. Kozak,V0120 + BL12 | ||
{|border="1" cellpadding="5" cellspacing="0" align="left" | |||
|- | |||
! scope="col" style="background:#efefef;" | # | |||
! scope="col" style="background:#efefef;" | Biobrick | |||
! scope="col" style="background:#efefef;" | ng/µL | |||
|- | |||
|1 | |||
|hPCD : mCh : SP1AB (BL01) | |||
|18.834 | |||
|- | |||
|2 | |||
|fshPCD : mCh : SP1AB (BL05) | |||
|14.471 | |||
|- | |||
|3 | |||
|flyPCD : mCh : SP1A (BL09) | |||
|7.302 | |||
|- | |||
|4 | |||
|hPCD : BL01 (BL12) | |||
|10.84 | |||
|- | |||
|}<br><br><br><br><br><br><br><br> | |||
{| {{table}} | |||
|align="center" style="background:#f0f0f1;"|'''µL of insert = (length insert/lengthvector *2*25 ng vector/concentration insert)''' | |||
|} | |||
{| {{table}} | {| {{table}} | ||
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* Meanwhile, get ice bucket and allow competent E. coli cells to thaw for five minutes. | * Meanwhile, get ice bucket and allow competent E. coli cells to thaw for five minutes. | ||
* Then follow transformation process: | * Then follow transformation process: | ||
'''Transformation Process''' | |||
*Warm five 100 μg/mL Amp agar plates at 37 °C | |||
*Thaw fresh tube of DH5α Turbo cells on ice | |||
*Add 30 μL of DH5α Turbo cells to DNA + dH2O | |||
*Include #5 tube, water only, no DNA (negative control) | |||
*Incubate cells + DNA on ice for 10 min. | |||
*Label pre-warmed plates | |||
*Transfer cells + DNA onto agar | |||
*Add 10 - 15 sterile glass beads, shake, discard beads | |||
*Incubate plates at 37 °C overnight | |||
[[Image:plates3_27_2013.jpg|200px|thumb|left|Ligation attempt 3/27/2013]] | |||
*''No colonies on any of the plates. Try again.'' | |||
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__NOTOC__ | __NOTOC__ |
Revision as of 18:21, 6 September 2013
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3/27/2013
Transformation Process
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