Description: 

This kit is designed to clean up and concentrate DNA from ChIP Assay or other samples containing small amount of DNA. With advance designed Tini Spin Column, it provides a hassle-free method for the rapid purification and concentration of high quality DNA from any standard ChIP protocol, including samples from: A) reverse cross-link, Proteinase K or RNase A digestion, mechanical or nudease-mediated DNA shearing and B) elution from chromatin-antibody-bead complexes in TES, 0.1M NaHCO3 and 1% SDS. The specially formulated ChIP DNA binding Buffer promotes the DNA binding to the column and purify the DNA from buffers containing detergents up to 5% Triton X-100, 5%Tween-20, 5% Sarkosyl, and 1%SDS. The kit can also be used to purify DNA or Oligo (17bp -10kb) from other enzymatic reactions. The elution volume of Tini spin column is as low as 5l, which can be used as a concentrator for DNA samples. 

Feature: 

- Easy and rapid with 10 min. procedure using Tini Spin Column. - High quality DNA recovery and concentration 

- Cleanup DNA from standard ChIP assay, mini prep, and enzymatic reactions (eg. labeling, restriction, and dephosphorlation) 

-Concentrate DNA to as low as 5l volume. –Recover DNA from 17bp-10kb 

 *Add 100% isopropanol before use: add 18 ml 100% isopropanol to12 ml ChIP DNA Binding Buffer. 

**Add 100% ethanol before use: add 24ml 100% ethanol to 6ml 5xWash Buffer. 

**Tini Spin columns can be order separately for leftover solutions (cat#EZC106, $39 for 50 columns) 

Caution: 

ChIP DNA Binding Buffer contains chaotropic salt. Please use proper safety precautions and always wear gloves when handling the reagent. Avoid contact with skin, eyes or clothing. In case of accidental spill or contact, wash thoroughly with water, seek medical advice if necessary. 

Procedures: 

1. Mix 5 volumes of the ChIP DNA Binding Buffer with each volume of sample (5:1). Mix briefly. 

Example a): Add 250l of the binding buffer to 50l cell lysate following DNA shearing, reverse cross-link and Proteinase K digestion in TES (50mM Tris,pH8.0, 10mM EDTA, 1% SDS) or 0.1M NaHCO3 containing 1% SDS. 

Example b): Add 500l of the binding buffer to 100l eluent in TES or 0.1M NaHCO3 containing 1% SDS buffers from chromatin-antibody-protein A agarose bead complex followed by reverse cross-link and Proteinase K digestion. 

Note: For cleanup DNA fragments <100bp, use 10 volumes of ChIP DNA Binding Buffer instead. 

2. Add 2.5l of 3M Sodium Acetate to a) or 5l of 3M Sodium Acetate to b) from step 1 and mix well. 

Note: For cleanup DNA and Oligo from other enzymatic reaction, it is not necessary to add Sodium Acetate. 

3. Load the sample mixture to Tini Spin Column (with collection tube) and spin in a microcentrifuge for 1 min at full speed (about 13,000 rpm). Do not load more than 350 l of sample on Tini spin column at one time. Discard the flow through and load more sample mixture if needed. 

4. Add 250l of Wash Buffer (Ethanol added) and centrifuge for 1 min. Repeat this step one more time (optional). 

5. Discard flow through and centrifuge at full speed with the lid open for 2 minutes to remove the ethanol completely. It is important to remove residual ethanol which may affect downstream applications. 

6. Add 5-50 l distilled water or Elution Buffer (preheated at 65oC for better yield) to the center of the column and leave at room temperature for 5 min. Spin the column for 1 min to elute the DNA from the column.