GDSYZX-United/NOTEBOOK/day note

Day note

2016.7.31

9:00

Extract genome from Arabidopsis Thaliana

The genome has been degraded greatly,possibly because we didn’t grind the leaves in very low temperature to inhibit the nuclease.We have to do it again.

2016.7.31

13:30

Extract genome from Arabidopsis Thaliana

We extracted it the genome successfully.But the genome has still been degraded a little. (Since we don’t have liquid nitrogen in our lab,so we grinded the leaves in an ice box. )We will use the genome to do the following PCR.

2016.8.1

9:00

We use PCR to generate and to amplify our target DNA fragments:promotercop1,promoter phyB,promoterpif1,gene hhl1,gene pHhl,gene flu.

We failed to get all our target DNA fragments .We will check the primers and the PCR program ,and then do it again.

2016.8.2

9:00

Do the PCR again to get our target DNA fragments.

We PCR gene pHhl1,gene hhl1,gene flu successfully. We will PCR cop1 together with pif1,phyB tomorrow.

2016.8.3

9:00

We use PCR to get cop1,phyB and pif1

In the first PCR we get cop 1 and pif1,we get higher concentration of phyB in the second time PCR.

2016.8.3

13:30

We use a kit box to purify our PCR products,so we can get our genes.

We did it.

2016.8.3

in the afternoon

We cut our DNA fragments using restriction enzymes:BsaⅠ,PstⅠ, EcorⅠ.After this process ,the gene fragments will have one overhang at both ends.The overhangs will help them splice other fragments.

We did it.