Serialization experiments

Summary

It is a complementary experiment to experimental protocols A and B of the differential display technique (DD) experiment and the gene expression series analysis experiment.

Modern Neuroscience Research Techniques

Author(s): U. Windhorst & H. Johansson Translated by Z. Q. Zhao Jun Chen

Operation method

Serialization experiments

Materials and Instruments

Solution and Buffer LoTE cDNA Label Ligase

Move

Experimental Programs A and B

1. Cut and purified double tags were joined in tandem: the

2. Incubate for 2 h at 16t℃.

3. Sample the double-labeled tandem onto one lane of an 8% poly(acrylamide) gel and electrophorese for 3 h at room temperature, 80°C, using a 100bpladder as a molecular weight standard.

4. EB staining.

5. Cut off the gel region containing 400bp or larger tandem.

6. Purify the DNA from the excised gel fragments to isolate the double-tagged fragments as described in protocol 8. However, do not incubate at 37°C but at 65°C and do not use a dry ice/ethanol ice bath before centrifugation.

7. After precipitation, rinse the precipitate twice with 70% ethanol, dry and redissolve in 40ul of LoTE.

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