Haynes Lab:Notebook/HPK-CFP insertion into Gal4EED/Luc using CRISPR/2015/04/15: Difference between revisions

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I'm going to set up two cultures for the two colonies on the sample plate, but am not very confident of the success of the ligation given the colonies on the negative control.
I'm going to set up two cultures for the two colonies on the sample plate, but am not very confident of the success of the ligation given the colonies on the negative control.


==Plasmid Miniprep of mCherry Vector==
<b>--DNA Concentration--</b>
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Sample ID'''
| align="center" style="background:#f0f0f0;"|'''Concentration (ng/µL)'''
| align="center" style="background:#f0f0f0;"|'''St Dev (ng/µL)'''
|-
| mCh Vector 1||392.1||15.5
|-
| mCh Vector 2||315.5||18.1
|-
| mCh Vector 3||319.7||6.3
|-
| mCh Vector 4||375.1||7.1
|}
-----
<b>--Restriction Digest--</b>
{| {{table}}
| align="center" style="background:#f0f0f0;"|'''Reagent'''
| align="center" style="background:#f0f0f0;"|'''Volume'''
|-
| DNA||600ng (1.9 - 1.5 µL)
|-
| 10x FastDigest Buffer||3
|-
| EcoRI||1
|-
| PstI||1
|-
| Water||23
|-
| Total||30
|}


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Revision as of 15:48, 15 April 2015

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Transformation Results for mCherry vector + gBlock

Plate # colonies
positive control many dozens
sample 2
negative control 2

I'm going to set up two cultures for the two colonies on the sample plate, but am not very confident of the success of the ligation given the colonies on the negative control.

Plasmid Miniprep of mCherry Vector

--DNA Concentration--

Sample ID Concentration (ng/µL) St Dev (ng/µL)
mCh Vector 1 392.1 15.5
mCh Vector 2 315.5 18.1
mCh Vector 3 319.7 6.3
mCh Vector 4 375.1 7.1

--Restriction Digest--

Reagent Volume
DNA 600ng (1.9 - 1.5 µL)
10x FastDigest Buffer 3
EcoRI 1
PstI 1
Water 23
Total 30


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