GE 100 reactions 79770 Benutzerhandbuch

22

spaced, a compression artifact is indicated. Try using the dITP reaction
mixture or a formamide gel.

Bands in 2 or 3 lanes
1. Heterogeneous template DNA (2 bands) caused by spontaneous deletions

arising during M13 phage growth. Try control DNA and limit phage growth to
less than 6-8 hours.

2. Insufficient mixing of reaction mixtures.
3. The sequence may be prone to compression artifacts in the gel.

Compressions occur when the DNA (usually G-C rich) synthesized by the
DNA polymerase does not remain fully denatured during electrophoresis. Try
using the dITP reaction mixture, or a 30-40% formamide gel.

If problems persist please contact USB Technical Support for assistance at
(800) 321-9322 or techsupport

@

usbweb.com in the United States. For your

authorized distributor and support staff outside the United States, contact your
local 

GE Healthcare

 office. Contact information is listed in the

back of this protocol booklet.

The control DNA included in the kit is from pUC18, a double-stranded circular
DNA of 2.7kb. A partial sequence of this DNA is given below (14).

(Universal cycle primer)

5'-G TTTTCCCAGT CACGACGTTG TA->

AACGCCAGGG TTTTCCCAGT CACGACGTTG TAAAACGACG GCCAGTGCCA

10

20

30

40

50

AGCTTGCATG CCTGCAGGTC GACTCTAGAG GATCCCCGGG TACCGAGCTC

60

70

80

90

100

GAATTCGTAA TCATGTCATA GCTGTTTCCT GTGTGAAATT GTTATCCGCT

<--CTTTAA CAATAGGCGA

110

120

130

140

150

CACAATTCCA CACAACATAC GAGCCGGAAG CATAAAGTGT AAAGCCTGGG

GTGTT-5'(Reverse cycle primer)

160

170

180

190

200

GTGCCTAATG AGTGAGCTAA CTCACATTAA TTGCGTTGCG CTCACTGCCC

210

220

230

240

250

GCTTTCCAGT CGGGAAACCT GTCGTGCCAG CTGCATTAAT GAATCGGCCA

260

270

280

290

300

ACGCGCGGGG AGAGGCGGTT TGCGTATTGG GCGCTCTTCC GCTTCCTCGC

310

320

330

340

350

TCACTGACTC GCTGCGCTCG GTCGTTCGGC TGCGGCGAGC GGTATCAGCT

360

370

380

390

400

CACTCAAAGG CGGTAATACG GTTATCCACA GAATCAGGGG ATAACGCAGC