Difference between revisions of "User:Alexander Boehme/Notebook/BioE140L"

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(Project Description/Abstract)
(Notes)
Line 37: Line 37:
 
* Place some notes here for visitors
 
* Place some notes here for visitors
  
* Example: This project is currently on hold until further notice.
+
38 uL ddH2O
 
+
5 ul 10x expand buffer
 +
5 ul 2mM dNTPs
 +
1 ul oligo mixture (100uM total, mixture of oligos after combination of 100uM stocks)
 +
0.75 ul Expand polymerase
 +
Program (can run JCA/PCA1)
 +
2 min initial denature at 94oC
 +
30 sec denature at 94oC
 +
30 sec anneal at 55oC [This should be the overlap temp of your oligos - vary as needed]
 +
30 sec extension at 72oC
 +
repeat 2-4 30 times total
  
 
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Revision as of 10:10, 12 March 2013

<!-- sibboleth --><div id="lncal1" style="border:0px;"><div style="display:none;" id="id">lncal1</div><div style="display:none;" id="dtext"></div><div style="display:none;" id="page">User:Alexander Boehme/Notebook/BioE140L</div><div style="display:none;" id="fmt">yyyy/MM/dd</div><div style="display:none;" id="css">OWWNB</div><div style="display:none;" id="month"></div><div style="display:none;" id="year"></div><div style="display:none;" id="readonly">Y</div></div>

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AsbF-2 Synthon Construction File


Mix oligios for asbF-2 synthon, do PCA1 protocol (514 bp, pca1)

PCR pca1 with asbF2-NewForward and asbF2-NewReverse by PCA2 protocol (468 bp, pca2)

Digest pca2 with EcoRI/BglII (yes, BglII), gp (452+11+5 bp, pcr_dig)

(get pre-digested vector from JCA of pBca9145-Bca1144 (EcoRI/BamHI, vect_dig)

Ligate pcr_dig and vect_dig, transform, pick white colonies, map, sequence

asbF2-NewForward ccaaaGAATTCCGTCTCATAGAAACCCATCC

asbF2-NewReverse gactgAGATCTCGTCTCAGGTCTCAGGATCCAG

Notes

  • Place some notes here for visitors

38 uL ddH2O 5 ul 10x expand buffer 5 ul 2mM dNTPs 1 ul oligo mixture (100uM total, mixture of oligos after combination of 100uM stocks) 0.75 ul Expand polymerase Program (can run JCA/PCA1) 2 min initial denature at 94oC 30 sec denature at 94oC 30 sec anneal at 55oC [This should be the overlap temp of your oligos - vary as needed] 30 sec extension at 72oC repeat 2-4 30 times total

Recent changes

18 October 2017

     15:56 

BME100 f2017:Group8 W1030 L4‎‎ (2 changes | history) . . (+426). . [Jennifer L. Brodsky‎; Jacob Tomas Harrison Haye‎]

     

15:56

(cur | prev) . . (+409). . Jennifer L. Brodsky (talk | contribs) (Research and Development)

     

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(cur | prev) . . (+17). . Jacob Tomas Harrison Haye (talk | contribs) (SNP Information & Primer Design)

     15:20  BME100 f2017:Group7 W0800 L4‎ (diff | hist) . . (-4). . Miguel R. Almanza Lopez (talk | contribs) (OUR TEAM)

     15:16 

(Upload log). .

[Miguel R. Almanza Lopez‎; Emily A. Hanzlick‎; Teleah Hancer‎ (2×); Kaifu Chen‎ (2×); Jacob Tomas Harrison Haye‎ (4×); Alivia Ankrum‎ (8×)]

     

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. . Miguel R. Almanza Lopez (talk | contribs) uploaded File:MralPoopy.jpg

     

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. . Kaifu Chen (talk | contribs) uploaded File:Jcm.jpg

     

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. . Alivia Ankrum (talk | contribs) uploaded a new version of File:Screen Shot 2017-10-18 at 1.26.01 PM.png

     

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. . Alivia Ankrum (talk | contribs) uploaded File:Screen Shot 2017-10-18 at 1.26.01 PM.png

     

13:28

. . Emily A. Hanzlick (talk | contribs) uploaded File:PolymearseChainReactionExtraCreditImageEAH.png

     

13:25

. . Teleah Hancer (talk | contribs) uploaded a new version of File:Teleah6.png

     

13:24

. . Teleah Hancer (talk | contribs) uploaded File:Teleah6.png

     14:40 

BME100 f2017:Group1 W1030 L4‎‎ (17 changes | history) . . (+1,361). . [Abby E. Krell‎ (7×); Emily A. Hanzlick‎ (10×)]

     

14:40

(cur | prev) . . (+33). . Abby E. Krell (talk | contribs) (Protocol)

     

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(cur | prev) . . (+178). . Abby E. Krell (talk | contribs) (Protocol)

     

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(cur | prev) . . (+4). . Abby E. Krell (talk | contribs) (Research and Development)

     

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(cur | prev) . . (0). . Abby E. Krell (talk | contribs) (Research and Development)

     

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(cur | prev) . . (+17). . Abby E. Krell (talk | contribs) (Protocol)

     

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(cur | prev) . . (+23). . Emily A. Hanzlick (talk | contribs) (Research and Development)

     

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(cur | prev) . . (-12). . Emily A. Hanzlick (talk | contribs) (Research and Development)

     

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(cur | prev) . . (+5). . Emily A. Hanzlick (talk | contribs) (Research and Development)

     

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(cur | prev) . . (+177). . Emily A. Hanzlick (talk | contribs) (Research and Development)

     

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(cur | prev) . . (+427). . Emily A. Hanzlick (talk | contribs) (Research and Development)

     

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(cur | prev) . . (+153). . Emily A. Hanzlick (talk | contribs) (Research and Development)

     

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(cur | prev) . . (+63). . Emily A. Hanzlick (talk | contribs) (Research and Development)

     

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(cur | prev) . . (+39). . Emily A. Hanzlick (talk | contribs) (Research and Development)

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BME100 f2017:Group12 W1030 L4‎‎ (8 changes | history) . . (+713). . [Alivia Ankrum‎ (8×)]

     

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(cur | prev) . . (0). . Alivia Ankrum (talk | contribs) (SNP Information & Primer Design)

     

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(cur | prev) . . (+30). . Alivia Ankrum (talk | contribs) (SNP Information & Primer Design)

     

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(cur | prev) . . (0). . Alivia Ankrum (talk | contribs) (SNP Information & Primer Design)

     

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(cur | prev) . . (+38). . Alivia Ankrum (talk | contribs) (SNP Information & Primer Design)

     

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(cur | prev) . . (+30). . Alivia Ankrum (talk | contribs) (SNP Information & Primer Design)

     

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(cur | prev) . . (+53). . Alivia Ankrum (talk | contribs) (SNP Information & Primer Design)

     

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(cur | prev) . . (+8). . Alivia Ankrum (talk | contribs) (SNP Information & Primer Design)

     

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(cur | prev) . . (+554). . Alivia Ankrum (talk | contribs) (SNP Information & Primer Design)

     13:57 

BME100 f2017:Group11 W1030 L4‎‎ (4 changes | history) . . (-98). . [Teleah Hancer‎; Zoe Marmitt‎ (3×)]

     

13:57

(cur | prev) . . (-1). . Zoe Marmitt (talk | contribs) (Protocol)

     

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(cur | prev) . . (+2). . Zoe Marmitt (talk | contribs) (Protocol)

     

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(cur | prev) . . (-108). . Zoe Marmitt (talk | contribs) (Protocol)

     

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(cur | prev) . . (+9). . Teleah Hancer (talk | contribs) (OUR TEAM)