Testing Charm DNA Kit on LCM Samples

Protocol Link

5/14/24 - Extracting DNA from LCM-captured cells (LCM date 5/8/24)

Used Charm Biotech Just-a-Tube ™ Laser Captured Microdissection (LCM) Sample Total DNA Purification Kit, protocol

Sectioning performed on 5/6/24 and LCM on 5/9/24. See details here

Protocol notes:

• Followed the Charm FFPE protocol almost exactly, but did all incubations at 56 ºC based on other Paxgene extraction protocols. Skipped the 94ºC incubation before the addition of ProK. Tried two different incubation times post ProK addition (in the incubator genie):
  • LCM sample #7: 1 hour incubation
  • LCM sample #11: 3 hour incubation

Qubit Results

• Used High Sensitivity DNA Qubit Protocol
• All samples read twice, standard only read once

DNA Standards: 60.85 (S1) & 21384.80 (S2)

colony_id	DNA_QBIT_1	DNA_QBIT_2	DNA_QBIT_3	DNA_QBIT_AVG
LCM sample #7	0.143	0.139	0.165	0.149 ng/uL
LCM sample #11	0.113	0.098	0.100	0.104 ng/uL

DNA Quality Check: Tapestation

Nothing! Again!

Full results can be found here

DNA Quality Check: Gel, using Kristina’s gel protocol at the bottom of this page.

Loaded 10 uL of extracted DNA into each well (3&4) – no DNA present.