Daniel:Notebook/HiResChrPaint/2013-4-26: Difference between revisions

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==Lambda Exonuclease Digest==
==Lambda Exonuclease Digest==
1. Prepared samples according to following table
{| class="wikitable" <hiddentext>generated with [[:de:Wikipedia:Helferlein/VBA-Macro for EXCEL tableconversion]] V1.8</hiddentext>
|- style="font-size:12pt" align="center"
| width="65" height="33" | Sample
| width="65" | ng/uL
| width="65" | uL sample
| width="65" | uL 10x buffer
| width="65" | uL Lambda Exo
| width="65" | uL ddH20
| width="65" | Total
|- style="font-size:12pt" align="center"
| height="15" | V4S3A
| align="center" | 102.2
| align="center" | 30
| align="center" | 5
| align="center" | 2
| align="center" | 13
| align="center" | 50
|- style="font-size:12pt" align="center"
| height="15" | V4S3B
| align="center" | 86.8
| align="center" | 30
| align="center" | 5
| align="center" | 2
| align="center" | 13
| align="center" | 50
|- style="font-size:12pt" align="center"
| height="15" | V6S3A
| align="center" | 80.1
| align="center" | 30
| align="center" | 5
| align="center" | 2
| align="center" | 13
| align="center" | 50
|- style="font-size:12pt" align="center"
| height="15" | V6S3B
| align="center" | 77.9
| align="center" | 30
| align="center" | 5
| align="center" | 2
| align="center" | 13
| align="center" | 50
|}
2. Incubate for 1 hour at 37 C
3. Heat shock 75 C 10 minutes
4. Ethanol Precipitation
==Ethanol Precipitation==
1. Add 250 uL nuclease free H20 to mixture (total 300 uL)
2. Add 900 uL 100% EtOH, 30 uL NaOAc, 2 uL glycoblue (2/300X)
3. Incubate overnight at -80C

Revision as of 19:11, 26 April 2013

Probe Prep Test (Started 4/22/2013)

Back to calendar

Lambda Exonuclease Digest

1. Prepared samples according to following table
Sample ng/uL uL sample uL 10x buffer uL Lambda Exo uL ddH20 Total
V4S3A 102.2 30 5 2 13 50
V4S3B 86.8 30 5 2 13 50
V6S3A 80.1 30 5 2 13 50
V6S3B 77.9 30 5 2 13 50
2. Incubate for 1 hour at 37 C
3. Heat shock 75 C 10 minutes
4. Ethanol Precipitation

Ethanol Precipitation

1. Add 250 uL nuclease free H20 to mixture (total 300 uL)
2. Add 900 uL 100% EtOH, 30 uL NaOAc, 2 uL glycoblue (2/300X)
3. Incubate overnight at -80C