Rui:LabNotes/SingleCell/2011-1-25: Difference between revisions
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100pg (1,2,3,5,8), M, 500pg (1,2,5,6,7), M | 100pg (1,2,3,5,8), M, 500pg (1,2,5,6,7), M | ||
[[File:Example.jpg]] | [[File:Example.jpg]] | ||
** Unspecific amplification from NTC, size range from 100-400bp | |||
** Size shift from NTC to 500pg was observed | |||
==PCR purification== | ==PCR purification== |
Revision as of 19:18, 24 February 2011
PCR result
QPCR curve
1. Empty lane w/o tube
2. ES-500pg (4); EB-500pg (4)
3. ES-100pg (4); EB-100pg (4)
4. ES-10pg (4); EB-10pg (4)
5. NTC (4); NTC (4)
TBE PAGE gel
*4ul PCR product + 2ul loading dye *Low Mass ladder *6% TBE-PAGE gel, running at 200V for 30min *Stain w/ SYBR Gold for 3 min
M, NTC (1,2,5,6,7), M, 10pg (1,2,5,6,7)
100pg (1,2,3,5,8), M, 500pg (1,2,5,6,7), M
** Unspecific amplification from NTC, size range from 100-400bp ** Size shift from NTC to 500pg was observed
PCR purification
*Combine 4 tubes for each sample *Add 16ul NaOAc and 400ul EtOH (R.T.), mix *Incubate -80C for 1 hour *Centrifuge at 10,000rpm 4C 30min **observed a huge white pellet in each tube **Qiagen PCR purification kit should be used as mentioned in original protocol *Wash with 70% EtOH (R.T.) *Resolve in 36ul Nf water, store at -80C