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參數(shù)資料
| 型號： | S7804 |
| 廠商： | Electronic Theatre Controls, Inc. |
| 英文描述： | CpG WIZ Prader-Willi/Angelman Amplification Kit |
| 中文描述： | 中央人民政府武進Prader-Willi/Angelman擴增試劑盒 |
| 文件頁數(shù)： | 26/32頁 |
| 文件大?。?/td> | 386K |
| 代理商： | S7804 |
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c. If performing "hot start" PCR using a "hot start" enzyme, verify the initial
denaturation/activation time of 12 minutes at 95°C.
d. Confirm that the PCR polymerase is still active.
e. Confirm that no additional Mg
2+
was added to the PCR reaction mix.
f. The optimal annealing temperature is 62°C. If items #a-e have not remedied
the problem, re-optimize annealing conditions to suit your amplification
instrument.
No amplification product is generated in the experimental
samples using the P and M primer sets, but products of the
correct size are observed with the control samples.
Potential Problem #1: Experimental DNA samples were degraded
prior to chemical modification.
Recommendation:
Purify the genomic DNA again and repeat the chemical modification.
Potential Problem #2: Chemically modified experimental DNA
samples were stored for more than two months prior to PCR.
Recommendation:
Repeat the chemical modification on new genomic DNA samples.
P or M primer sets are producing bands in all samples,
including the "no DNA" controls.
Potential Problem: PCR reagents are contaminated with
amplification products.
Recommendations: see Sec. VI.
Appendix, Laboratory Setup and Precautions
a. Use fresh aliquots of every PCR component (i.e. dNTPs, buffer, etc.)
b. Use separate sets of pipettors for pre- vs. post-amplification liquid
dispensing.
c. Devote a work area to pre- and post-amplification procedures.
d. Always use aerosol-resistant pipette tips.
e. Always use a clean labcoat and gloves.
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