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From 2010.igem.org

Contents 1 Week #4 27th September - 01 October 2010 1.1 Monday 1.1.1 Well we are stuck with the vector this week we have to get vector is our main objetive. 1.2 Tuesday 1.2.1 We run with a low meelting's agarose prove to extract band tomorrow 1.2.2 we do a protocol to precipit with alchol an salts our objetive is do it all 1.2.3 to get vector and do the ligations. 1.3 Wensday 1.3.1 We prepared all to do ligations PsB1C3 with our Pcr's 1.3.2 for this we going to cut with EcoRI and PstI both vector and insert 1.3.3 We run a low melting gel slow 2 hours and a half then purified from the band

Week #4 27th September - 01 October 2010

Monday

Well we are stuck with the vector this week we have to get vector is our main objetive.

• We have achived a nanodrop readings as below

	ng/μl	260/280	260/230
PsB1C3	0.60	-0.73	-0.0
PCR 1 red	427.70	1.71	1.71
PCR 2 red	483.20	1.74	1.71
PCR 3 red	410.70	1.76	2.02
PCR 4 red	431.05	1.61	1.88
PCR 1 blue	533.35	1.71	1.75
PCR 2 blue	536.00	1.72	1.82
PCR 3 blue	577.50	1.69	1.59
PCR 4 blue	627.55	1.70	1.56
PsB1C3	4.10	2.62	1.01

• Yep our trouble is the lisis alcaline method to do the mini prep

we have a low concentration of vector’s plasmid DNA.

We have to work an try to get more plasmid and make dilutions of PCR’s

is not posible to do ligations with this ratio betwen vector an insert.

On this step we advisor has proposed a method via Low Meelting agarose

to geting out plasmid from the band.

Tuesday

We run with a low meelting's agarose prove to extract band tomorrow

we do a protocol to precipit with alchol an salts our objetive is do it all

to get vector and do the ligations.

Wensday

We prepared all to do ligations PsB1C3 with our Pcr's

for this we going to cut with EcoRI and PstI both vector and insert

We run a low melting gel slow 2 hours and a half then purified from the band