12/09/13

From 2013.igem.org

(Difference between revisions)
(→Gel Purification of the pGEM-T pasmid double digestions ran in the 11/09/13)
(→Double Digest)
Line 71: Line 71:
|}
|}
-
==Double Digest==
+
==Digestion of the samples purified with the enzyme PstI-HF==
 +
 
 +
{|border=1
 +
|'''DNA volume'''||'''Volume'''||'''Cutsmart buffer'''||'''Enzyme volume'''||'''dH2O added to total volume of 60 ul'''
 +
|-
 +
|TODX||23|6||1||30
 +
|-
 +
|TODF||22.5||6||1||30.5
 +
|-
 +
TOBG||6||1||31
 +
|-
 +
|pSB1C3 (RFP)||20.5||6||1||32.5
 +
|}
[[File:doubledigest_gelpurification_12092013.jpg]]
[[File:doubledigest_gelpurification_12092013.jpg]]

Revision as of 20:26, 13 September 2013

Home Team Official Team Profile Project Parts Submitted to the Registry Modeling Notebook Safety Attributions


Contents

Isolate plasmid from overnight culture

  • Using Omega mini prep kit protocol
  • Samples:
    • TODF 1,2,3,
    • TODX 1, 2, 3
    • TOBG 1, 2, 3,
    • RFP 1, 2, 3
sampleVolume (ul)Concentration (ng/ul)
TODF143120
TODF24398.8
TODX142399.3
TODX244442.2
TODX343417.5
TOBG143262.4
TOBG247319.2
TOBG341338.4
RFP14177.7
RFP24132.7
RFP34259

Glycerol stocks

  • Took 750ul from overnight culture and centrifuged at full speed for ~2 minutes
  • Supernatant was removed
  • Pellet resuspended in 375ul of HMFM
  • Samples were then frozen at -80

Gel Purification of the pGEM-T pasmid double digestions ran in the 11/09/13

The ge was purified by using the The Zymoclean Gel DNA recovery Kit and its protocol was followed.

Nanodrop Results

SampleVolumeConcentration (ng/ul)280/260260/230
TODX48.57.11.830.72
TODF45.521.71.891.19
pSB1C3 (RFP)46.27.31.810.16

Digestion of the samples purified with the enzyme PstI-HF

TOBG||6||1||31
DNA volumeVolumeCutsmart bufferEnzyme volumedH2O added to total volume of 60 ul
TODX6130
TODF22.56130.5
pSB1C3 (RFP)20.56132.5

Doubledigest gelpurification 12092013.jpg

Retrieved from "http://2013.igem.org/12/09/13"