Navigation Links
InstaGene Matrix: Prepare DNA Templates for PCR With No Phenol/Chloroform Extractions and No Deproteinization Steps, Rev C


Prepare DNA from blood, cultured cells, or bacteria:
Just incubate sample with InstaGene matrix at 56C for 1530 min, then boil for 8 min.

InstaGene matrix absorbs cell lysis products, producing an improved substrate for PCR amplification.

DNA Preparation From Whole Blood
Use whole blood samples that are frozen, refrigerated, or collected fresh.
1. Add 36 l of whole blood to 1 ml of autoclaved double distilled water in a 1.5 ml microfuge tube. Mix by inverting the tube several times.

2. Incubate the tube at room temperature for 1530 min.

3. Spin at 10,00012,000 rpm for 23 min.

4. Carefully remove all but 2030 l of the supernatant. Do not disturb the pellet.

5. Add 200 l of InstaGene matrix to the pellet and incubate at 56C for 1530 min.

Note: InstaGene matrix should be mixed at moderate speed on a magnetic stirrer to maintain the matrix in suspension. The pipet tip used should have a large bore, such as on a 1,000 l pipet tip (Bio-Rads TBR tips, catalog #223-9350 and 223-9351).

6. Vortex at high speed for 10 sec. Place the tube in a 100C heating block or boiling water bath for 8 min.

7. Vortex at high speed for 10 sec. Spin at 10,00012,000 rpm for 23 min.

8. Use 20 l of the resulting supernatant per 50 l PCR reaction. Store the remainder at -20C. Repeat step 7 when reusing the InstaGene DNA preparation.

Note: It is important to store the prepared sample at -20C when not in use.

DNA Preparation From Cultured Mammalian Cells
1. Pellet 200 l of cells suspension from media in a microfuge tube. Spin at 10,00012,000 rpm for 1 min and remove the supernatant.

2. Resuspend cells in 1 ml of 1x PBS and centrifuge for 1 min at 10,00012,000 rpm.

3. Resuspend cells in autoclaved water at 2030 cells/l.

4. Add 20 l of this cell suspension to 200 l of InstaGene matrix. Incubate at 56C for 1530 min.

Note: InstaGene matrix should be mixed at moderate speed on a magnetic stirrer to maintain the matrix in suspension. The pipet tip used should have a large bore, such as on a 1,000 l pipet tip.

5. Follow the procedure for DNA preparation from whole blood, steps 68.

DNA Preparation From Bacteria
The protocol described below is for the preparation of genomic DNA or episomal DNA from bacteria.
1. Pick an isolated bacterial colony and resuspend it in 1 ml of autoclaved water in a microfuge tube.

2. Centrifuge for 1 min at 10,00012,000 rpm. Remove the supernatant.

3. Follow the DNA preparation procedure for whole blood, steps 58.


back to top
'"/>

Source:


Page: All 1 2

Related biology technology :

1. Preparation of PCR Quality Template DNA from a Wide Range of Tissues Using InstaGene Matrix
2. Prepare siRNA and miRNA Probes in Just 1 Hour
3. Rapidly Prepare Probes and Markers for Small RNA Analysis
4. TripleMaster and Perfect gDNA Blood Mini Kit Team Up to Amplify Long gDNA Templates
5. Using the TripleMaster PCR System for Robust Amplification of GC-Rich DNA Templates
6. Inactivation of Contaminating Templates in PCR Using High Intensity Ultraviolet Light
Post Your Comments:
*Name:
*Comment:
*Email:


(Date:10/12/2017)... ... 12, 2017 , ... AMRI, a global contract research, development ... patient outcomes and quality of life, will now be offering its impurity solutions ... new regulatory requirements for all new drug products, including the finalization of ICH ...
(Date:10/11/2017)... YORBA LINDA, CA (PRWEB) , ... October 11, ... ... adapted to upregulate any gene in its endogenous context, enabling overexpression experiments and ... activation (CRISPRa) system with small RNA guides is transformative for performing systematic gain-of-function ...
(Date:10/11/2017)... ... 2017 , ... Proscia Inc ., a data solutions ... “Pathology is going digital. Is your lab ready?” with Dr. Nicolas Cacciabeve, Managing ... how Proscia improves lab economics and realizes an increase in diagnostic confidence.* ...
(Date:10/11/2017)... ... October 11, 2017 , ... Disappearing forests and increased emissions ... over 5.5 million people each year. Especially those living in larger cities are affected ... based in one of the most pollution-affected countries globally - decided to take action. ...
Breaking Biology Technology:
(Date:4/5/2017)... , April 4, 2017 KEY FINDINGS ... anticipated to expand at a CAGR of 25.76% during ... diseases is the primary factor for the growth of ... report: https://www.reportbuyer.com/product/4807905/ MARKET INSIGHTS The global ... product, technology, application, and geography. The stem cell market ...
(Date:3/30/2017)... 30, 2017  On April 6-7, 2017, Sequencing.com will ... hackathon at Microsoft,s headquarters in ... focus on developing health and wellness apps that provide ... the Genome is the first hackathon for personal ... largest companies in the genomics, tech and health industries ...
(Date:3/29/2017)...  higi, the health IT company that operates the ... , today announced a Series B investment from ... The new investment and acquisition accelerates higi,s strategy to ... population health activities through the collection and workflow integration ... collects and secures data today on behalf of over ...
Breaking Biology News(10 mins):