Haynes Lab:Notebook:Shared/Cloning Bootcamp/2015/11/20: Difference between revisions

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==Baylee Murphy- 11/20/15==
==Baylee Murphy- 11/20/15 Session 3==


KAH126- MV2 Culture inoculation results from [http://openwetware.org/wiki/Haynes_Lab:Notebook:Shared/Cloning_Bootcamp/2015/11/18 Session 2]
KAH126- MV2 Culture inoculation results from [http://openwetware.org/wiki/Haynes_Lab:Notebook:Shared/Cloning_Bootcamp/2015/11/18 Session 2]
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Results from NanoDrop
Results from NanoDrop


'''Excel table'''
{|
| align="center" style="background:#f0f0f0;"|'''Sample Read#'''
| align="center" style="background:#f0f0f0;"|'''Location'''
| align="center" style="background:#f0f0f0;"|'''260 Raw'''
| align="center" style="background:#f0f0f0;"|'''280 Raw'''
| align="center" style="background:#f0f0f0;"|'''320 Raw'''
| align="center" style="background:#f0f0f0;"|'''260'''
| align="center" style="background:#f0f0f0;"|'''280'''
| align="center" style="background:#f0f0f0;"|'''260/280'''
| align="center" style="background:#f0f0f0;"|'''ng/µL'''
|-
| BM KAH126-1||C2||0.32||0.207||0.08||0.249||0.131||1.909||249.344
|-
| BM KAH126-1||C3||0.313||0.201||0.074||0.245||0.128||1.911||244.882
|-
| BM KAH126-2||D2||0.323||0.203||0.06||0.272||0.146||1.861||272.416
|-
| BM KAH126-2||D3||0.319||0.2||0.066||0.258||0.136||1.904||258.244
|}




Using Xba restriction enzyme which will preform 2 cuts- expending bands at 1934 bp and 4505 bp
Using Xba restriction enzyme which will preform 2 cuts- expending bands at 1934 bp and 4505 bp


'''insert picture'''
[[Image:Screen Shot 2015-11-21 at 4.30.33 PM.png]]


Results of restriction digest show expected bands and indicate Xba enzyme worked




Revision as of 16:38, 21 November 2015

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Baylee Murphy- 11/20/15 Session 3

KAH126- MV2 Culture inoculation results from Session 2

  • Liquid cultures 1 and 2 both had cloudy yellow broth, indicating growth
  • Streaks for both showed colony growth

Samples for mini prep and analysis

  • (1) Sample 1- KAH126-MV2 colony 1, [BB#}; part = [length in bp]; vector= [length in bp]
  • (2) Sample 2- KAH126-MV2 colony 2, [BB#}; part = [length in bp]; vector= [length in bp]

Results from NanoDrop

Sample Read# Location 260 Raw 280 Raw 320 Raw 260 280 260/280 ng/µL
BM KAH126-1 C2 0.32 0.207 0.08 0.249 0.131 1.909 249.344
BM KAH126-1 C3 0.313 0.201 0.074 0.245 0.128 1.911 244.882
BM KAH126-2 D2 0.323 0.203 0.06 0.272 0.146 1.861 272.416
BM KAH126-2 D3 0.319 0.2 0.066 0.258 0.136 1.904 258.244


Using Xba restriction enzyme which will preform 2 cuts- expending bands at 1934 bp and 4505 bp

Results of restriction digest show expected bands and indicate Xba enzyme worked


plasmid validation template page

KAH126 incluswa Hpdc, scar, mcherry: about 1000 bp

  • forward primer- DD122 10 uM
  • reverse primer-DD123 10 uM


Samples sent for sequencing at Bio Design around 4:45pm

  • (1) Sample 1- KAH126-MV2 colony 1, forward primer DD122
  • (2) Sample 2- KAH126-MV2 colony 1, reverse primer DD123
  • (3) Sample 3- KAH126-MV2 colony 2, forward primer DD122
  • (4) Sample 4- KAH126-MV2 colony 2, reverse primer DD123






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