DNeasy Blood & Tissue Kit

DNA isolation / purification Cells - primary cells mouse embryonic fibroblast (MEF)

Experiment
DNA isolation / purification Cells - primary cells mouse embryonic fibroblast (MEF)
Product
DNeasy Blood & Tissue Kit from Qiagen
Manufacturer
Qiagen

Protocol tips

Upstream tips
DO NOT add bleach or acidic solutions directly to the sample-preparation
waste.
Downstream tips
- Include RNAse treatment for 15-20 min.
- Ensure EtOH is completely evaporated off of the column prior to elution. Adjust time from 1min to 5 min at 60`C
- Use prewarmed TE buffer to elute the DNA

Publication protocol

Telomere length
Telomere length was determined via the SYBR green qPCR procedure described by Cawthorn et al. [18]. Briefly, telomere length is proportional to the ratio between the telomere Ct value and a single copy gene Ct value using the DNA primers listed in table 1. Genomic DNA was extracted using the DNeasy blood and tissue kit (69504, Qiagen). A master mix with 20 ng of the template genomic DNA, 10 µM forward primer, 10 µM reverse primer, and fast SYBR green master mix (4346906, Applied Biosystems) for the telomere and single copy gene primers. Identical plates were set up for each primer set. The Applied Biosystems 7900HT Fast Real-Time PCR System was used for detection with the temperature profile listed in Table 2. Ct values were automatically generated with the RQ Manager Version 1.2 qPCR software (Applied Biosystems).

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Discussion

Discussion

1 year ago

Author: R. Verma India

DNA isolation column clogged

During centrifugation, the column got clogged and I was unable to continue with the protocol. How can I unclog it?

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Papers

Check out relevant papers found by Labettor's AI that are relevant for performing DNA isolation / purification Cells - primary cells mouse embryonic fibroblast (MEF) using DNeasy Blood & Tissue Kit from Qiagen.

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Manufacturer protocol

Download the product protocol from Qiagen for DNeasy Blood & Tissue Kit below.

Download PDF Download manufacturer protocol

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